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Sample records for buds

  1. Bud Dormancy and Growth

    Science.gov (United States)

    Nearly all land plants produce ancillary meristems in the form of axillary or adventitious buds in addition to the shoot apical meristem. Outgrowth of these buds has a significant impact on plant architecture and the ability of plants to compete with neighboring plants, as well as to respond to and ...

  2. Tumor Budding in Colorectal Carcinomas

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    Sevda SERT BEKTAŞ

    2012-01-01

    Full Text Available Objective: In colorectal carcinomas, tumor budding has been defined as the presence of isolated single tumor cells or small cell clusters in the stroma at the invasive tumor margin. In this study, the relationship between tumor budding density at the invasive tumor margin and pathological parameters is investigated.Material and Method: Haematoxylin and eosin stained slides of 73 cases with colorectal carcinoma were retrospectively evaluated for the presence and intensity of tumor budding by 2 observers. After the specimens were assessed, the highest density of tumor budding area was counted in a microscopic field of x200. Cases were separated into 2 groups according to tumor budding density as low grade (<10 and high grade (≥10. The relationship of these groups with depth of tumor invasion, histological grade, vascular invasion and lymph node involvement was investigated.Results: Of the 73 colorectal carcinoma cases, 33 (45.2% had low and 40 (54.8% had high grade tumor budding density, respectively. There was a statistically significant relationship between high grade tumor budding density and histological grade (p=0.042, lymph node involvement (p=0.0001 and vascular invasion (p=0.0034.Conclusion: High grade tumor budding density is associated with aggressive phenotypical features in colorectal carcinoma.

  3. Budding of Walnut ( Juglans regia L.

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    Majlind Kasmi

    2013-09-01

    Full Text Available The walnut is classified as a strategic species for human nutrition and is included in the FAO’s list of priority plants. Walnut, (Juglans regia L. propagation is more difficult, compared to most fruit species. Due to walnut heterozygosity, propagation by seeds does not lead to inheritance of all the characteristics of certain varieties. That is the reason why propagation technologies are being improved worldwide. The purpose of this experiment was to increase the success of inoculation of the walnut budding var. Franquete. Methods such as the patch budding and chip budding have been employed during the experiment. To establish the most appropriate season of inoculation, June budding on 28 June (with buds taken in the current season, autumn budding on 28 August (with buds taken in the current season and spring budding on 28 May (with buds collected from the winter dormant period, were tested. As rootstocks for the June and August budding, the seedlings of Juglans regia L. of the current year's growth have been employed. For the spring inoculation the one year old scions have been used. Patch budding resulted the most successful method for walnuts. However, the success of the method of patch budding depends on the season of inoculation. An 80 % of successful inoculation was achieved by June budding (on 28 June. Furthermore, cutting off the leaf 20 days before the buds being taken for budding, led to even higher results reaching 87% of successful inoculation. According to the results of the present study, the June budding of the patch method seems to be the best solution for the production of grafted young walnut trees.

  4. Tropical Storms Bud and Dera

    Science.gov (United States)

    2002-01-01

    Like dancers pirouetting in opposite directions, the rotational patterns of two different tropical storms are contrasted in this pair of Multi-angle Imaging Spectroradiometer (MISR) nadir-camera images. The left-hand image is of Tropical Storm Bud, acquired on June 17, 2000 (Terra orbit 2656) as the storm was dissipating. Bud was situated in the eastern Pacific Ocean between Socorro Island and the southern tip of Baja California. South of the storm's center is a vortex pattern caused by obstruction of the prevailing flow by tiny Socorro Island. Sonora, Mexico and Baja California are visible at the top of the image. The right-hand image is of Tropical Cyclone Dera, acquired on March 12, 2001. Dera was located in the Indian Ocean, south of Madagascar. The southern end of this large island is visible in the top portion of this image. Northern hemisphere tropical storms, like Bud, rotate in a counterclockwise direction, whereas those in the southern hemisphere, such as Dera, rotate clockwise. The opposite spins are a consequence of Earth's rotation. Each image covers a swath approximately 380 kilometers wide. Image courtesy NASA/JPL/GSFC/LaRC, MISR Team

  5. Whole-Transcriptome Analysis of Differentially Expressed Genes in the Vegetative Buds, Floral Buds and Buds of Chrysanthemum morifolium.

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    Hua Liu

    Full Text Available Chrysanthemum morifolium is an important floral crop that is cultivated worldwide. However, due to a lack of genomic resources, very little information is available concerning the molecular mechanisms of flower development in chrysanthemum.The transcriptomes of chrysanthemum vegetative buds, floral buds and buds were sequenced using Illumina paired-end sequencing technology. A total of 15.4 Gb of reads were assembled into 91,367 unigenes with an average length of 739 bp. A total of 43,137 unigenes showed similarity to known proteins in the Swissprot or NCBI non-redundant protein databases. Additionally, 25,424, 24,321 and 13,704 unigenes were assigned to 56 gene ontology (GO categories, 25 EuKaryotic Orthologous Groups (KOG categories, and 285 Kyoto Encyclopedia of Genes and Genomes (KEGG pathways, respectively. A total of 1,876 differentially expressed genes (DEGs (1,516 up-regulated, 360 down-regulated were identified between vegetative buds and floral buds, and 3,300 DEGs (1,277 up-regulated, 1,706 down-regulated were identified between floral buds and buds. Many genes encoding important transcription factors (e.g., AP2, MYB, MYC, WRKY, NAC and CRT as well as proteins involved in carbohydrate metabolism, protein kinase activity, plant hormone signal transduction, and the defense responses, among others, were considerably up-regulated in floral buds. Genes involved in the photoperiod pathway and flower organ determination were also identified. These genes represent important candidate genes for molecular cloning and functional analysis to study flowering regulation in chrysanthemum.This comparative transcriptome analysis revealed significant differences in gene expression and signaling pathway components between the vegetative buds, floral buds and buds of Chrysanthemum morifolium. A wide range of genes was implicated in regulating the phase transition from vegetative to reproductive growth. These results should aid researchers in the study of

  6. HIV Pol inhibits HIV budding and mediates the severe budding defect of Gag-Pol.

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    Xin Gan

    Full Text Available The prevailing hypothesis of HIV budding posits that the viral Gag protein drives budding, and that the Gag p6 peptide plays an essential role by recruiting host-cell budding factors to sites of HIV assembly. HIV also expresses a second Gag protein, p160 Gag-Pol, which lacks p6 and fails to bud from cells, consistent with the prevailing hypothesis of HIV budding. However, we show here that the severe budding defect of Gag-Pol is not caused by the absence of p6, but rather, by the presence of Pol. Specifically, we show that (i the budding defect of Gag-Pol is unaffected by loss of HIV protease activity and is therefore an intrinsic property of the Gag-Pol polyprotein, (ii the N-terminal 433 amino acids of Gag and Gag-Pol are sufficient to drive virus budding even though they lack p6, (iii the severe budding defect of Gag-Pol is caused by a dominant, cis-acting inhibitor of budding in the HIV Pol domain, and (iv Gag-Pol inhibits Gag and virus budding in trans, even at normal levels of Gag and Gag-Pol expression. These and other data support an alternative hypothesis of HIV budding as a process that is mediated by the normal, non-viral pathway of exosome/microvesicle biogenesis.

  7. Repellence of the red bud borer (Resseliella oculiperda) to grafted apple trees by impregnation of budding tape with essential oils

    NARCIS (Netherlands)

    Tol, van R.W.H.M.; Linden, van der A.; Swarts, H.J.; Visser, J.H.

    2007-01-01

    The red bud borer Resseliella oculiperda (Rübs.) is a pest insect of apple trees when rootstocks are grafted with scion buds by shield budding. The female midges are attracted to the wounds of the grafted buds where they lay their eggs. The larvae feed on the cambium and destroy the buds completely

  8. Shrinkage of ipsilateral taste buds and hyperplasia of contralateral taste buds following chorda tympani nerve transection

    Institute of Scientific and Technical Information of China (English)

    Yi-ke Li; Juan-mei Yang; Yi-bo Huang; Dong-dong Ren; Fang-lu Chi

    2015-01-01

    The morphological changes that occur in the taste buds after denervation are not well under-stood in rats, especially in the contralateral tongue epithelium. In this study, we investigated the time course of morphological changes in the taste buds following unilateral nerve transection. The role of the trigeminal component of the lingual nerve in maintaining the structural integrity of the taste buds was also examined. Twenty-four Sprague-Dawley rats were randomly divided into three groups:control, unilateral chorda tympani nerve transection and unilateral chorda tympani nerve transection+lingual nerve transection. Rats were allowed up to 42 days of re-covery before being euthanized. The taste buds were visualized using a cytokeratin 8 antibody. Taste bud counts, volumes and taste receptor cell numbers were quantiifed and compared among groups. No signiifcant difference was detected between the chorda tympani nerve transection and chorda tympani nerve transection+lingual nerve transection groups. Taste bud counts, vol-umes and taste receptor cell numbers on the ipsilateral side all decreased signiifcantly compared with control. On the contralateral side, the number of taste buds remained unchanged over time, but they were larger, and taste receptor cells were more numerous postoperatively. There was no evidence for a role of the trigeminal branch of the lingual nerve in maintaining the structural integrity of the anterior taste buds.

  9. Shrinkage of ipsilateral taste buds and hyperplasia of contralateral taste buds following chorda tympani nerve transection

    Directory of Open Access Journals (Sweden)

    Yi-ke Li

    2015-01-01

    Full Text Available The morphological changes that occur in the taste buds after denervation are not well understood in rats, especially in the contralateral tongue epithelium. In this study, we investigated the time course of morphological changes in the taste buds following unilateral nerve transection. The role of the trigeminal component of the lingual nerve in maintaining the structural integrity of the taste buds was also examined. Twenty-four Sprague-Dawley rats were randomly divided into three groups: control, unilateral chorda tympani nerve transection and unilateral chorda tympani nerve transection + lingual nerve transection. Rats were allowed up to 42 days of recovery before being euthanized. The taste buds were visualized using a cytokeratin 8 antibody. Taste bud counts, volumes and taste receptor cell numbers were quantified and compared among groups. No significant difference was detected between the chorda tympani nerve transection and chorda tympani nerve transection + lingual nerve transection groups. Taste bud counts, volumes and taste receptor cell numbers on the ipsilateral side all decreased significantly compared with control. On the contralateral side, the number of taste buds remained unchanged over time, but they were larger, and taste receptor cells were more numerous postoperatively. There was no evidence for a role of the trigeminal branch of the lingual nerve in maintaining the structural integrity of the anterior taste buds.

  10. Oxytocin signaling in mouse taste buds.

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    Michael S Sinclair

    Full Text Available BACKGROUND: The neuropeptide, oxytocin (OXT, acts on brain circuits to inhibit food intake. Mutant mice lacking OXT (OXT knockout overconsume salty and sweet (i.e. sucrose, saccharin solutions. We asked if OXT might also act on taste buds via its receptor, OXTR. METHODOLOGY/PRINCIPAL FINDINGS: Using RT-PCR, we detected the expression of OXTR in taste buds throughout the oral cavity, but not in adjacent non-taste lingual epithelium. By immunostaining tissues from OXTR-YFP knock-in mice, we found that OXTR is expressed in a subset of Glial-like (Type I taste cells, and also in cells on the periphery of taste buds. Single-cell RT-PCR confirmed this cell-type assignment. Using Ca2+ imaging, we observed that physiologically appropriate concentrations of OXT evoked [Ca2+]i mobilization in a subset of taste cells (EC50 approximately 33 nM. OXT-evoked responses were significantly inhibited by the OXTR antagonist, L-371,257. Isolated OXT-responsive taste cells were neither Receptor (Type II nor Presynaptic (Type III cells, consistent with our immunofluorescence observations. We also investigated the source of OXT peptide that may act on taste cells. Both RT-PCR and immunostaining suggest that the OXT peptide is not produced in taste buds or in their associated nerves. Finally, we also examined the morphology of taste buds from mice that lack OXTR. Taste buds and their constituent cell types appeared very similar in mice with two, one or no copies of the OXTR gene. CONCLUSIONS/SIGNIFICANCE: We conclude that OXT elicits Ca2+ signals via OXTR in murine taste buds. OXT-responsive cells are most likely a subset of Glial-like (Type I taste cells. OXT itself is not produced locally in taste tissue and is likely delivered through the circulation. Loss of OXTR does not grossly alter the morphology of any of the cell types contained in taste buds. Instead, we speculate that OXT-responsive Glial-like (Type I taste bud cells modulate taste signaling and afferent

  11. Competitive canalization of PIN-dependent auxin flow from axillary buds controls pea bud outgrowth.

    Science.gov (United States)

    Balla, Jozef; Kalousek, Petr; Reinöhl, Vilém; Friml, Jiří; Procházka, Stanislav

    2011-02-01

    Shoot branching is one of the major determinants of plant architecture. Polar auxin transport in stems is necessary for the control of bud outgrowth by a dominant apex. Here, we show that following decapitation in pea (Pisum sativum L.), the axillary buds establish directional auxin export by subcellular polarization of PIN auxin transporters. Apical auxin application on the decapitated stem prevents this PIN polarization and canalization of laterally applied auxin. These results support a model in which the apical and lateral auxin sources compete for primary channels of auxin transport in the stem to control the outgrowth of axillary buds. PMID:21219506

  12. Identification and Quality Assessment of Chrysanthemum Buds by CE Fingerprinting

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    Xiaoping Xing

    2015-01-01

    Full Text Available A simple and efficient fingerprinting method for chrysanthemum buds was developed with the aim of establishing a quality control protocol based on biochemical makeup. Chrysanthemum bud samples were successively extracted by water and alcohol. The fingerprints of the chrysanthemum buds samples were obtained using capillary electrophoresis and electrochemical detection (CE-ED employing copper and carbon working electrodes to capture all of the chemical information. 10 batches of chrysanthemum buds were collected from different regions and various factories to establish the baseline fingerprint. The experimental data of 10 batches electropherogram buds by CE were analyzed by correlation coefficient and the included angle cosine methods. A standard chrysanthemum bud fingerprint including 24 common peaks was established, 12 from each electrode, which was successfully applied to identify and distinguish between chrysanthemum buds from 2 other chrysanthemum species. These results demonstrate that fingerprint analysis can be used as an important criterion for chrysanthemum buds quality control.

  13. Bilingual Buds: The Evolution of a Program

    Science.gov (United States)

    Huang, Sharon

    2009-01-01

    The impetus to begin Bilingual Buds came about six years ago when the author, pregnant with twins and commuting into New York City, was reading about the numerous cognitive benefits for children of acquiring a second language early in their lives. She was surprised to learn that even by the age of six months, children begin to lose the ability to…

  14. Bipolar budding in yeasts - an electron microscope study

    NARCIS (Netherlands)

    Kreger-van Rij, N.J.W.; Veenhuis, M.

    1971-01-01

    Bud formation in yeasts with bipolar budding was studied by electron microscopy of thin sections. Budding in yeasts of the species Saccharomycodes ludwigii, Hanseniaspora valbyensis and Wickerhamia fluorescens resulted in concentric rings of scar ridges on the wall of the mother cell. The wall betwe

  15. Synchronization of the Budding Yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Foltman, Magdalena; Molist, Iago; Sanchez-Diaz, Alberto

    2016-01-01

    A number of model organisms have provided the basis for our understanding of the eukaryotic cell cycle. These model organisms are generally much easier to manipulate than mammalian cells and as such provide amenable tools for extensive genetic and biochemical analysis. One of the most common model organisms used to study the cell cycle is the budding yeast Saccharomyces cerevisiae. This model provides the ability to synchronise cells efficiently at different stages of the cell cycle, which in turn opens up the possibility for extensive and detailed study of mechanisms regulating the eukaryotic cell cycle. Here, we describe methods in which budding yeast cells are arrested at a particular phase of the cell cycle and then released from the block, permitting the study of molecular mechanisms that drive the progression through the cell cycle.

  16. Molecular Mechanism of Arenavirus Assembly and Budding

    OpenAIRE

    Shuzo Urata; Jiro Yasuda

    2012-01-01

    Arenaviruses have a bisegmented negative-strand RNA genome, which encodes four viral proteins: GP and NP by the S segment and L and Z by the L segment. These four viral proteins possess multiple functions in infection, replication and release of progeny viruses from infected cells. The small RING finger protein, Z protein is a matrix protein that plays a central role in viral assembly and budding. Although all arenaviruses encode Z protein, amino acid sequence alignment showed a huge variety ...

  17. Mechanism of supercooling in flower bud of Camellia oleifea

    Institute of Scientific and Technical Information of China (English)

    苏维埃; 潘良文

    1995-01-01

    It is the first time for MRI to be used in the research of flower buds supercooling. Directobservation on freezing course of living flower buds of Camellia yuhsienensis by MRI and tissue browning test showed that freezing order of the flower organs is bud axis, scale, petal, pistil and stamen. It is coincident with the direction of ice development from bud axes to flower organs upwards. The corresponding results from MRI and freezing-fixation showed that the water translocation from flower organs to axes and scales is carried on in the course of bud freezing. ’H spectral measurement of NMR was used to follow the decrease of unfrozen water in the buds during the cooling.

  18. Breadth of Tuning and Taste Coding in Mammalian Taste Buds

    OpenAIRE

    Tomchik, Seth M.; Berg, Stephanie; Kim, Joung Woul; Chaudhari, Nirupa; Roper, Stephen D.

    2007-01-01

    A longstanding question in taste research concerns taste coding and, in particular, how broadly are individual taste bud cells tuned to taste qualities (sweet, bitter, umami, salty, and sour). Taste bud cells express G-protein-coupled receptors for sweet, bitter, or umami tastes but not in combination. However, responses to multiple taste qualities have been recorded in individual taste cells. We and others have shown previously there are two classes of taste bud cells directly involved in gu...

  19. A Monitor for Bud Emergence in the Yeast Morphogenesis Checkpoint

    OpenAIRE

    Theesfeld, Chandra L.; Zyla, Trevin R.; Bardes, Elaine G.S.; Lew, Daniel J.

    2003-01-01

    Cell cycle transitions are subject to regulation by both external signals and internal checkpoints that monitor satisfactory progression of key cell cycle events. In budding yeast, the morphogenesis checkpoint arrests the cell cycle in response to perturbations that affect the actin cytoskeleton and bud formation. Herein, we identify a step in this checkpoint pathway that seems to be directly responsive to bud emergence. Activation of the kinase Hsl1p is dependent upon...

  20. Processing Umami and Other Tastes in Mammalian Taste Buds

    OpenAIRE

    Roper, Stephen D.; Chaudhari, Nirupa

    2009-01-01

    Neuroscientists are now coming to appreciate that a significant degree of information processing occurs in the peripheral sensory organs of taste prior to signals propagating to the brain. Gustatory stimulation causes taste bud cells to secrete neurotransmitters that act on adjacent taste bud cells (paracrine transmitters) as well as on primary sensory afferent fibers (neurocrine transmitters). Paracrine transmission, representing cell-cell communication within the taste bud, has the potentia...

  1. Essential Oil of Betula pendula Roth. Buds

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    Betül Demirci

    2004-01-01

    Full Text Available The essential oil of Betula pendula Roth. buds was obtained using both hydrodistillation and microdistillation techniques and their chemical compositions were analyzed using both gas chromatography (GC and gas chromatography–mass spectrometry (GC-MS. Overall, more than 50 compounds were identified representing 80% and 92% for hydrodistillation and microdistillation, respectively. The main components (by hydrodistillation and microdistillation, respectively found were α-copaene (12% and 10%, germacrene D (11% and 18% and δ-cadinene (11% and 15% in the analyzed essential oils. The microdistillation technique proved to be a useful tool and compliant alternative when compared to hydrodistillation.

  2. Taxonomy Icon Data: Budding yeast [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available Budding yeast Saccharomyces cerevisiae Saccharomyces_cerevisiae_L.png Saccharomyces..._cerevisiae_NL.png Saccharomyces_cerevisiae_S.png Saccharomyces_cerevisiae_NS.png http://biosciencedbc.jp/taxonomy..._icon/icon.cgi?i=Saccharomyces+cerevisiae&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Saccharomy...ces+cerevisiae&t=NL http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Saccharomy...ces+cerevisiae&t=S http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Saccharomyces+cerevisiae&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=216 ...

  3. A quantitative study of cat epiglottal taste buds during development.

    OpenAIRE

    Stedman, H M; MISTRETTA, C.M.; Bradley, R M

    1983-01-01

    To quantify the development of upper airway chemoreceptors, epiglottal taste buds were counted in cat fetuses, kittens and adults. Taste buds with structural characteristics similar to those in the adult were observed in all fetuses close to term and in kittens from birth. During the first postnatal week a mean of 76 taste buds was present on the kitten epiglottis and by adulthood 800 were observed. The number of taste buds increased as a logarithmic function of both age and weight, but the c...

  4. Development Correlations of the Buds of Grapevine (Vitis vinifera L.

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    Liliana ROTARU

    2010-06-01

    Full Text Available The development characteristics of different buds of the grapevine are mainly related by stimulation and/or inhibition effects, the action of which is still inexplicable. The present study examines the development dynamics of the buds of a one-year old branch after excision of different buds and the application of ?-naphtyl acetic acid (ANA, as well as the growth capacity of each bud individually. We verified the effects of acrotony cited previously by various researchers. These effects are due to different developmental characteristics of which could to lay the groundwork for the improvement of different productions methods.

  5. Tumor budding in upper gastrointestinal carcinomas

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    Viktor Hendrik Koelzer

    2014-08-01

    Full Text Available The basis of personalized medicine in oncology is the prediction of an individual’s risk of relapse and death from disease. The presence of tumor budding (TB at the tumor-host interface of gastrointestinal cancers has been recognized as a hallmark of unfavorable disease biology. TB is defined as the presence of dedifferentiated cells or small clusters of up to five cells at the tumor invasive front and can be observed in aggressive carcinomas of the esophagus, stomach, pancreas, ampulla, colon and rectum. Presence of TB reproducibly correlates with advanced tumor stage, frequent lymphovascular invasion, nodal and distant metastasis. The UICC has officially recognized TB as additional independent prognostic factor in cancers of the colon and rectum. Recent studies have also characterized TB as a promising prognostic indicator for clinical management of esophageal squamous cell carcinoma, adenocarcinoma of the gastro-esophageal junction and gastric adenocarcinoma. However, several important issues have to be addressed for application in daily diagnostic practice: 1 Validation of prognostic scoring systems for tumor budding in large, multi-center studies 2 Consensus on the optimal assessment method 3 Inter-observer reproducibility. This review provides a comprehensive analysis of TB in cancers of the upper gastrointestinal tract including critical appraisal of perspectives for further study.

  6. Specific residues of the GDP/GTP exchange factor Bud5p are involved in establishment of the cell type-specific budding pattern in yeast.

    Science.gov (United States)

    Kang, Pil Jung; Lee, Bongyong; Park, Hay-Oak

    2004-07-01

    Cells of the budding yeast undergo oriented cell division by choosing a specific site for growth depending on their cell type. Haploid a and alpha cells bud in an axial pattern whereas diploid a/alpha cells bud in a bipolar pattern. The Ras-like GTPase Rsr1p/Bud1p, its GDP-GTP exchange factor Bud5p, and its GTPase-activating protein Bud2p are essential for selecting the proper site for polarized growth in all cell types. Here we showed that specific residues at the N terminus and the C terminus of Bud5p were important for bipolar budding, while some residues were involved in both axial and bipolar budding. These bipolar-specific mutations of BUD5 disrupted proper localization of Bud5p in diploid a/alpha cells without affecting Bud5p localization in haploid alpha cells. In contrast, Bud5p expressed in the bud5 mutants defective in both budding patterns failed to localize in all cell types. Thus, these results identify specific residues of Bud5p that are likely to be involved in direct interaction with spatial landmarks, which recruit Bud5p to the proper bud site. Finally, we found a new start codon of BUD5, which extends the open reading frame to 210 bp upstream of the previously estimated start site, thus encoding a polypeptide of 608 amino acid residues. Bud5p with these additional N-terminal residues interacted with Bud8p, a potential bipolar landmark, suggesting that the N-terminal region is necessary for recognition of the spatial cues. PMID:15136576

  7. Photosynthetic leaf area modulates tiller bud outgrowth in sorghum.

    Science.gov (United States)

    Kebrom, Tesfamichael H; Mullet, John E

    2015-08-01

    Shoot branches or tillers develop from axillary buds. The dormancy versus outgrowth fates of buds depends on genetic, environmental and hormonal signals. Defoliation inhibits bud outgrowth indicating the role of leaf-derived metabolic factors such as sucrose in bud outgrowth. In this study, the sensitivity of bud outgrowth to selective defoliation was investigated. At 6 d after planting (6 DAP), the first two leaves of sorghum were fully expanded and the third was partially emerged. Therefore, the leaves were selectively defoliated at 6 DAP and the length of the bud in the first leaf axil was measured at 8 DAP. Bud outgrowth was inhibited by defoliation of only 2 cm from the tip of the second leaf blade. The expression of dormancy and sucrose-starvation marker genes was up-regulated and cell cycle and sucrose-inducible genes was down-regulated during the first 24 h post-defoliation of the second leaf. At 48 h, the expression of these genes was similar to controls as the defoliated plant recovers. Our results demonstrate that small changes in photosynthetic leaf area affect the propensity of tiller buds for outgrowth. Therefore, variation in leaf area and photosynthetic activity should be included when integrating sucrose into models of shoot branching.

  8. A Monitor for Bud Emergence in the Yeast Morphogenesis Checkpoint

    Science.gov (United States)

    Theesfeld, Chandra L.; Zyla, Trevin R.; Bardes, Elaine G.S.; Lew, Daniel J.

    2003-01-01

    Cell cycle transitions are subject to regulation by both external signals and internal checkpoints that monitor satisfactory progression of key cell cycle events. In budding yeast, the morphogenesis checkpoint arrests the cell cycle in response to perturbations that affect the actin cytoskeleton and bud formation. Herein, we identify a step in this checkpoint pathway that seems to be directly responsive to bud emergence. Activation of the kinase Hsl1p is dependent upon its recruitment to a cortical domain organized by the septins, a family of conserved filament-forming proteins. Under conditions that delayed or blocked bud emergence, Hsl1p recruitment to the septin cortex still took place, but hyperphosphorylation of Hsl1p and recruitment of the Hsl1p-binding protein Hsl7p to the septin cortex only occurred after bud emergence. At this time, the septin cortex spread to form a collar between mother and bud, and Hsl1p and Hsl7p were restricted to the bud side of the septin collar. We discuss models for translating cellular geometry (in this case, the emergence of a bud) into biochemical signals regulating cell proliferation. PMID:12925763

  9. 5'-end sequences of budding yeast full-length cDNA clones - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available Budding yeast cDNA sequencing project 5'-end sequences of budding yeast full-length cDNA clones Data detail Data name 5'-end sequence...s of budding yeast full-length cDNA clones Description of data contents cDNA sequence...e Update History of This Database Site Policy | Contact Us 5'-end sequences of budding yeast full-length cDNA clones - Budding yeast cDNA sequencing project | LSDB Archive ...

  10. Budding yeast for budding geneticists: a primer on the Saccharomyces cerevisiae model system.

    Science.gov (United States)

    Duina, Andrea A; Miller, Mary E; Keeney, Jill B

    2014-05-01

    The budding yeast Saccharomyces cerevisiae is a powerful model organism for studying fundamental aspects of eukaryotic cell biology. This Primer article presents a brief historical perspective on the emergence of this organism as a premier experimental system over the course of the past century. An overview of the central features of the S. cerevisiae genome, including the nature of its genetic elements and general organization, is also provided. Some of the most common experimental tools and resources available to yeast geneticists are presented in a way designed to engage and challenge undergraduate and graduate students eager to learn more about the experimental amenability of budding yeast. Finally, a discussion of several major discoveries derived from yeast studies highlights the far-reaching impact that the yeast system has had and will continue to have on our understanding of a variety of cellular processes relevant to all eukaryotes, including humans.

  11. This bud's for you: mechanisms of cellular nucleocytoplasmic trafficking via nuclear envelope budding.

    Science.gov (United States)

    Fradkin, Lee G; Budnik, Vivian

    2016-08-01

    The nuclear envelope (NE) physically separates the cytoplasmic and nuclear compartments. While this barrier provides advantages, it also presents a challenge for the nuclear export of large ribonucleoprotein (RNP) complexes. Decades-old dogma holds that all such border-crossing is via the nuclear pore complex (NPC). However, the diameter of the NPC central channel limits the passage of large cargos. Here, we review evidence that such large RNPs employ an endogenous NE-budding pathway, previously thought to be exclusive to the nuclear egress of Herpes viruses. We discuss this and other models proposed, the likelihood that this pathway is conserved, and the consequences of disrupting NE-budding for synapse development, localized translation of synaptic mRNAs, and laminopathies inducing accelerated aging. PMID:27236823

  12. Calling Card Analysis in Budding Yeast.

    Science.gov (United States)

    Mayhew, David; Mitra, Robi D

    2016-02-01

    Calling card analysis is a high-throughput method for identifying the genomic binding sites of multiple transcription factors in a single experiment in budding yeast. By tagging a DNA-binding protein with a targeting domain that directs the insertion of the Ty5 retrotransposon, the genomic binding sites for that transcription factor are marked. The transposition locations are then identified en masse by Illumina sequencing. The calling card protocol allows for simultaneous analysis of multiple transcription factors. By cloning barcodes into the Ty5 transposon, it is possible to pair a unique barcode with every transcription factor in the experiment. The method presented here uses expression of transcription factors from their native loci; however, it can also be altered to measure binding sites of transcription factors overexpressed from a plasmid. PMID:26832687

  13. Measuring mitotic spindle dynamics in budding yeast

    Science.gov (United States)

    Plumb, Kemp

    In order to carry out its life cycle and produce viable progeny through cell division, a cell must successfully coordinate and execute a number of complex processes with high fidelity, in an environment dominated by thermal noise. One important example of such a process is the assembly and positioning of the mitotic spindle prior to chromosome segregation. The mitotic spindle is a modular structure composed of two spindle pole bodies, separated in space and spanned by filamentous proteins called microtubules, along which the genetic material of the cell is held. The spindle is responsible for alignment and subsequent segregation of chromosomes into two equal parts; proper spindle positioning and timing ensure that genetic material is appropriately divided amongst mother and daughter cells. In this thesis, I describe fluorescence confocal microscopy and automated image analysis algorithms, which I have used to observe and analyze the real space dynamics of the mitotic spindle in budding yeast. The software can locate structures in three spatial dimensions and track their movement in time. By selecting fluorescent proteins which specifically label the spindle poles and cell periphery, mitotic spindle dynamics have been measured in a coordinate system relevant to the cell division. I describe how I have characterised the accuracy and precision of the algorithms by simulating fluorescence data for both spindle poles and the budding yeast cell surface. In this thesis I also describe the construction of a microfluidic apparatus that allows for the measurement of long time-scale dynamics of individual cells and the development of a cell population. The tools developed in this thesis work will facilitate in-depth quantitative analysis of the non-equilibrium processes in living cells.

  14. A permeability barrier surrounds taste buds in lingual epithelia.

    Science.gov (United States)

    Dando, Robin; Pereira, Elizabeth; Kurian, Mani; Barro-Soria, Rene; Chaudhari, Nirupa; Roper, Stephen D

    2015-01-01

    Epithelial tissues are characterized by specialized cell-cell junctions, typically localized to the apical regions of cells. These junctions are formed by interacting membrane proteins and by cytoskeletal and extracellular matrix components. Within the lingual epithelium, tight junctions join the apical tips of the gustatory sensory cells in taste buds. These junctions constitute a selective barrier that limits penetration of chemosensory stimuli into taste buds (Michlig et al. J Comp Neurol 502: 1003-1011, 2007). We tested the ability of chemical compounds to permeate into sensory end organs in the lingual epithelium. Our findings reveal a robust barrier that surrounds the entire body of taste buds, not limited to the apical tight junctions. This barrier prevents penetration of many, but not all, compounds, whether they are applied topically, injected into the parenchyma of the tongue, or circulating in the blood supply, into taste buds. Enzymatic treatments indicate that this barrier likely includes glycosaminoglycans, as it was disrupted by chondroitinase but, less effectively, by proteases. The barrier surrounding taste buds could also be disrupted by brief treatment of lingual tissue samples with DMSO. Brief exposure of lingual slices to DMSO did not affect the ability of taste buds within the slice to respond to chemical stimulation. The existence of a highly impermeable barrier surrounding taste buds and methods to break through this barrier may be relevant to basic research and to clinical treatments of taste.

  15. Taste bud homeostasis in health, disease, and aging.

    Science.gov (United States)

    Feng, Pu; Huang, Liquan; Wang, Hong

    2014-01-01

    The mammalian taste bud is an onion-shaped epithelial structure with 50-100 tightly packed cells, including taste receptor cells, supporting cells, and basal cells. Taste receptor cells detect nutrients and toxins in the oral cavity and transmit the sensory information to gustatory nerve endings in the buds. Supporting cells may play a role in the clearance of excess neurotransmitters after their release from taste receptor cells. Basal cells are precursor cells that differentiate into mature taste cells. Similar to other epithelial cells, taste cells turn over continuously, with an average life span of about 8-12 days. To maintain structural homeostasis in taste buds, new cells are generated to replace dying cells. Several recent studies using genetic lineage tracing methods have identified populations of progenitor/stem cells for taste buds, although contributions of these progenitor/stem cell populations to taste bud homeostasis have yet to be fully determined. Some regulatory factors of taste cell differentiation and degeneration have been identified, but our understanding of these aspects of taste bud homoeostasis remains limited. Many patients with various diseases develop taste disorders, including taste loss and taste distortion. Decline in taste function also occurs during aging. Recent studies suggest that disruption or alteration of taste bud homeostasis may contribute to taste dysfunction associated with disease and aging. PMID:24287552

  16. Characterization of Septin Ultrastructure in Budding Yeast Using Electron Tomography

    Science.gov (United States)

    Bertin, Aurélie; Nogales, Eva

    2015-01-01

    Summary Septins are essential for the completion of cytokinesis. In budding yeast, Saccharomyces cerevisiae, septins are located at the bud neck during mitosis and are closely connected to the inner plasma membrane. In vitro, yeast septins have been shown to self-assemble into a variety of filamentous structures, including rods, paired filaments, bundles and rings [1–3]. Using electron tomography of freeze-substituted section and cryo-electron tomography of frozen sections, we determined the three dimensional organization of the septin cytoskeleton in dividing budding yeast with molecular resolution [4,5]. Here we describe the detailed procedures used for our characterization of the septin cellular ultrastructure. PMID:26519309

  17. Mechanical feedback stabilizes budding yeast morphogenesis

    Science.gov (United States)

    Banavar, Samhita; Trogdon, Michael; Petzold, Linda; Campas, Otger

    Walled cells have the ability to remodel their shape while sustaining an internal turgor pressure that can reach values up to 10 atmospheres. This requires a tight and simultaneous regulation of cell wall assembly and mechanochemistry, but the underlying mechanisms by which this is achieved remain unclear. Using the growth of mating projections in budding yeast (S. cerevisiae) as a motivating example, we have developed a theoretical description that couples the mechanics of cell wall expansion and assembly via a mechanical feedback. In the absence of a mechanical feedback, cell morphogenesis is inherently unstable. The presence of a mechanical feedback stabilizes changes in cell shape and growth, and provides a mechanism to prevent cell lysis in a wide range of conditions. We solve for the dynamics of the system and obtain the different dynamical regimes. In particular, we show that several parameters affect the stability of growth, including the strength of mechanical feedback in the system. Finally, we compare our results to existing experimental data.

  18. Flower Bud Differentiation in Quercus suber L.

    Directory of Open Access Journals (Sweden)

    Maria Carolina Varela

    2016-06-01

    Full Text Available Background and Purpose: Cork oak (Quercus suber L. is one of the most important forest species growing in the Western Mediterranean region. This investigation intends to assess the timing of flowering differentiation of cork oak and contribute to the deepening of the knowledge about the process of the sexual reproduction of the species. Materials and Methods: In 2010 four trees were selected (9, 14, 24, 25 from a plot of 25 trees located at Quinta da Serra, Portugal. A total of 240 buds were collected from these four trees, on three days (8, 14 and 23 March, from 4 branches per tree and 5 positions per branch for the assessment of meristem differentiation. Results: Meristem differentiation analysed on the sampling days revealed there were only vegetative structures by 8 March; a few male and female primordia on 14 March; and fully differentiated reproductive structures on 23 March. Conclusions: Flowering sex determination of cork oak occurs about one month before the flowering onset.

  19. Apoptosis at inflection point in liquid culture of budding yeasts.

    Directory of Open Access Journals (Sweden)

    Toshiyuki Hagiwara

    Full Text Available Budding yeasts are highly suitable for aging studies, because the number of bud scars (stage proportionally correlates with age. Its maximum stages are known to reach at 20-30 stages on an isolated agar medium. However, their stage dynamics in a liquid culture is virtually unknown. We investigate the population dynamics by counting scars in each cell. Here one cell division produces one new cell and one bud scar. This simple rule leads to a conservation law: "The total number of bud scars is equal to the total number of cells." We find a large discrepancy: extremely fewer cells with over 5 scars than expected. Almost all cells with 6 or more scars disappear within a short period of time in the late log phase (corresponds to the inflection point. This discrepancy is confirmed directly by the microscopic observations of broken cells. This finding implies apoptosis in older cells (6 scars or more.

  20. A novel role for the GTPase-activating protein Bud2 in the spindle position checkpoint.

    Directory of Open Access Journals (Sweden)

    Scott A Nelson

    Full Text Available The spindle position checkpoint (SPC ensures correct mitotic spindle position before allowing mitotic exit in the budding yeast Saccharomyces cerevisiae. In a candidate screen for checkpoint genes, we identified bud2Δ as deficient for the SPC. Bud2 is a GTPase activating protein (GAP, and the only known substrate of Bud2 was Rsr1/Bud1, a Ras-like GTPase and a central component of the bud-site-selection pathway. Mutants lacking Rsr1/Bud1 had no checkpoint defect, as did strains lacking and overexpressing Bud5, a guanine-nucleotide exchange factor (GEF for Rsr1/Bud1. Thus, the checkpoint function of Bud2 is distinct from its role in bud site selection. The catalytic activity of the Bud2 GAP domain was required for the checkpoint, based on the failure of the known catalytic point mutant Bud2(R682A to function in the checkpoint. Based on assays of heterozygous diploids, bud2(R682A, was dominant for loss of checkpoint but recessive for bud-site-selection failure, further indicating a separation of function. Tem1 is a Ras-like protein and is the critical regulator of mitotic exit, sitting atop the mitotic exit network (MEN. Tem1 is a likely target for Bud2, supported by genetic analyses that exclude other Ras-like proteins.

  1. Functional cell types in taste buds have distinct longevities.

    Directory of Open Access Journals (Sweden)

    Isabel Perea-Martinez

    Full Text Available Taste buds are clusters of polarized sensory cells embedded in stratified oral epithelium. In adult mammals, taste buds turn over continuously and are replenished through the birth of new cells in the basal layer of the surrounding non-sensory epithelium. The half-life of cells in mammalian taste buds has been estimated as 8-12 days on average. Yet, earlier studies did not address whether the now well-defined functional taste bud cell types all exhibit the same lifetime. We employed a recently developed thymidine analog, 5-ethynil-2'-deoxyuridine (EdU to re-evaluate the incorporation of newly born cells into circumvallate taste buds of adult mice. By combining EdU-labeling with immunostaining for selected markers, we tracked the differentiation and lifespan of the constituent cell types of taste buds. EdU was primarily incorporated into basal extragemmal cells, the principal source for replenishing taste bud cells. Undifferentiated EdU-labeled cells began migrating into circumvallate taste buds within 1 day of their birth. Type II (Receptor taste cells began to differentiate from EdU-labeled precursors beginning 2 days after birth and then were eliminated with a half-life of 8 days. Type III (Presynaptic taste cells began differentiating after a delay of 3 days after EdU-labeling, and they survived much longer, with a half-life of 22 days. We also scored taste bud cells that belong to neither Type II nor Type III, a heterogeneous group that includes mostly Type I cells, and also undifferentiated or immature cells. A non-linear decay fit described these cells as two sub-populations with half-lives of 8 and 24 days respectively. Our data suggest that many post-mitotic cells may remain quiescent within taste buds before differentiating into mature taste cells. A small number of slow-cycling cells may also exist within the perimeter of the taste bud. Based on their incidence, we hypothesize that these may be progenitors for Type III cells.

  2. Real Life Science with Dandelions and Project BudBurst

    OpenAIRE

    Johnson, Katherine A.

    2016-01-01

    Project BudBurst is a national citizen-science project that tracks bloom times and other phenological data for plants across the country. Data from Project BudBurst are being used to measure the effects of climate change. Students can participate in this project by watching any of the plants on the list, including the common dandelion, which makes the program easy and accessible to everyone. Journal of Microbiology & Biology Education

  3. Taste Bud Homeostasis in Health, Disease, and Aging

    OpenAIRE

    Feng, Pu; Huang, Liquan; Wang, Hong

    2013-01-01

    The mammalian taste bud is an onion-shaped epithelial structure with 50–100 tightly packed cells, including taste receptor cells, supporting cells, and basal cells. Taste receptor cells detect nutrients and toxins in the oral cavity and transmit the sensory information to gustatory nerve endings in the buds. Supporting cells may play a role in the clearance of excess neurotransmitters after their release from taste receptor cells. Basal cells are precursor cells that differentiate into mature...

  4. A buckling mechanism for ESCRT-III budding

    CERN Document Server

    Lenz, Martin; Joanny, Jean-François

    2009-01-01

    The ESCRT-III protein complex binds to the membrane of eukaryotic cells, causing it to bud into long tubes. Here we propose that this budding is akin to a buckling instability. We analyze the linear stability of flat ESCRT-III-dressed membranes and account for the formation of long tubes. We study strongly deformed dressed membranes and their bifurcation diagram numerically. Our mechanism is compatible with reasonable in vivo parameter values and we propose an experiment allowing its validation.

  5. Dielectric modelling of cell division for budding and fission yeast

    International Nuclear Information System (INIS)

    The frequency dependence of complex permittivity or the dielectric spectrum of a system including a cell in cell division has been simulated by a numerical technique based on the three-dimensional finite difference method. Two different types of cell division characteristic of budding and fission yeast were examined. The yeast cells are both regarded as a body of rotation, and thus have anisotropic polarization, i.e. the effective permittivity of the cell depends on the orientation of the cell to the direction of an applied electric field. In the perpendicular orientation, where the rotational axis of the cell is perpendicular to the electric field direction, the dielectric spectra for both yeast cells included one dielectric relaxation and its intensity depended on the cell volume. In the parallel orientation, on the other hand, two dielectric relaxations appeared with bud growth for budding yeast and with septum formation for fission yeast. The low-frequency relaxation was shifted to a lower frequency region by narrowing the neck between the bud and the mother cell for budding yeast and by increasing the degree of septum formation for fission yeast. After cell separation, the low-frequency relaxation disappeared. The simulations well interpreted the oscillation of the relative permittivity of culture broth found for synchronous cell growth of budding yeast

  6. Dielectric modelling of cell division for budding and fission yeast

    Science.gov (United States)

    Asami, Koji; Sekine, Katsuhisa

    2007-02-01

    The frequency dependence of complex permittivity or the dielectric spectrum of a system including a cell in cell division has been simulated by a numerical technique based on the three-dimensional finite difference method. Two different types of cell division characteristic of budding and fission yeast were examined. The yeast cells are both regarded as a body of rotation, and thus have anisotropic polarization, i.e. the effective permittivity of the cell depends on the orientation of the cell to the direction of an applied electric field. In the perpendicular orientation, where the rotational axis of the cell is perpendicular to the electric field direction, the dielectric spectra for both yeast cells included one dielectric relaxation and its intensity depended on the cell volume. In the parallel orientation, on the other hand, two dielectric relaxations appeared with bud growth for budding yeast and with septum formation for fission yeast. The low-frequency relaxation was shifted to a lower frequency region by narrowing the neck between the bud and the mother cell for budding yeast and by increasing the degree of septum formation for fission yeast. After cell separation, the low-frequency relaxation disappeared. The simulations well interpreted the oscillation of the relative permittivity of culture broth found for synchronous cell growth of budding yeast.

  7. Tumor Budding: The Name is EMT. Partial EMT.

    Science.gov (United States)

    Grigore, Alexandru Dan; Jolly, Mohit Kumar; Jia, Dongya; Farach-Carson, Mary C; Levine, Herbert

    2016-01-01

    Tumor budding is a histological phenomenon encountered in various cancers, whereby individual malignant cells and/or small clusters of malignant cells are seen in the tumor stroma. Postulated to be mirror epithelial-mesenchymal transition, tumor budding has been associated with poor cancer outcomes. However, the vast heterogeneity in its exact definition, methodology of assessment, and patient stratification need to be resolved before it can be routinely used as a standardized prognostic feature. Here, we discuss the heterogeneity in defining and assessing tumor budding, its clinical significance across multiple cancer types, and its prospective implementation in clinical practice. Next, we review the emerging evidence about partial, rather than complete, epithelial-mesenchymal phenotype at the tumor bud level, and its connection with tumor proliferation, quiescence, and stemness. Finally, based on recent literature, indicating a co-expression of epithelial and mesenchymal markers in many tumor buds, we posit tumor budding to be a manifestation of this hybrid epithelial/mesenchymal phenotype displaying collective cell migration.

  8. Optimum harvesting time of herbaceous peony buds for cutting flowers

    Institute of Scientific and Technical Information of China (English)

    YU Xiao-nan; GUO Peng-peng; LU Guang-pei; ZHANG Qi-xiang

    2011-01-01

    The herbaceous peony is one of the cut flowers that has become increasingly popular in the international market in recent years. In the study,11 cultivars of herbaceous peonies suitable for cutting flowers were selected;different harvesting stages (three or four stages) were identified according to bud development observation (bud firmness, bud diameter, sepal angle, petal angle, and color showing). Moreover, flower development and vase life were also recorded in the experiment of vase. Bud development observation shows that there are great differences in optimum harvesting stage between the cultivars, especially between the hybrid peony group and lactiflora group. This implies that bud diameter could not be the only indicator for optimum harvesting stagc. Bud description, including sepal opening angle, sepal color, petal color showing, and degree of firmness, is necessary for the determination of the harvesting stage. Among the 11 cultivars, Pink Hawaiian Coral', ‘Red Charm', ‘Edulis Superba', ‘Red Magic', and ‘Sarah Bernhardt' should be harvested at stage 1. 'Duchesse de Nemours', Taff, ‘Sorbet' and ‘Monsieur Jules Elie' should be harvested at stage 2. ‘Kansas' should be at stage 3 and ‘Karl Rosenfield' at stage 4. The determination of optimum harvesting stage for each cultivar is an important element in cut flower production.

  9. [Bud population dynamics of Phragmites australis in heterogeneous habitats of Northeast grassland, China].

    Science.gov (United States)

    2015-02-01

    To adapt ecological environment, typical clonal plants can occur continuously by means of buds. The changes in the bud bank and bud flow in the heterogeneous habitats become the foundation for deep understanding the characteristics of vegetative propagation. By sampling soil from the unit area, a comparative analysis was performed for rhizome bud population dynamics of Phragmites australis community in both meadow soil and saline-alkali soil habitats in meadow grassland of Northeast China. The one-age class rhizome buds formed in the current year were used as input, with the other age classes rhizome buds as output, counting the dormancy buds and death buds. The results showed that the storage, input, output, dormancy, death and the input rates of P. australis rhizome bud populations in meadow soil habitat were significantly higher than that in saline-alkali habitat. There was no significant difference in output rate between the two habitats. The dormant rate in saline-alkali habitat was significantly greater than that in meadow soil habitat. The death rates remained at relatively low levels in both, less than 2%. With the going of growing season, the input buds and input rate of bud bank increased in the two habitats, while the output buds remained relatively stable. The output rate increased first and decreased later, the dormancy buds and dormant rate decreased. Bud bank and bud flow were positively related to soil moisture, soil organic matter and soil available nitrogen content. However, they were negatively related to soil pH value and soil available phosphorus content. Bud bank and bud flow had a similar seasonal variation. Constantly for both habitats, P. australis populations generated new rhizome buds supplied to the bud bank and kept a stable output to maintain their vegetative propagation.

  10. Electron tomography reveals the steps in filovirus budding.

    Directory of Open Access Journals (Sweden)

    Sonja Welsch

    2010-04-01

    Full Text Available The filoviruses, Marburg and Ebola, are non-segmented negative-strand RNA viruses causing severe hemorrhagic fever with high mortality rates in humans and nonhuman primates. The sequence of events that leads to release of filovirus particles from cells is poorly understood. Two contrasting mechanisms have been proposed, one proceeding via a "submarine-like" budding with the helical nucleocapsid emerging parallel to the plasma membrane, and the other via perpendicular "rocket-like" protrusion. Here we have infected cells with Marburg virus under BSL-4 containment conditions, and reconstructed the sequence of steps in the budding process in three dimensions using electron tomography of plastic-embedded cells. We find that highly infectious filamentous particles are released at early stages in infection. Budding proceeds via lateral association of intracellular nucleocapsid along its whole length with the plasma membrane, followed by rapid envelopment initiated at one end of the nucleocapsid, leading to a protruding intermediate. Scission results in local membrane instability at the rear of the virus. After prolonged infection, increased vesiculation of the plasma membrane correlates with changes in shape and infectivity of released viruses. Our observations demonstrate a cellular determinant of virus shape. They reconcile the contrasting models of filovirus budding and allow us to describe the sequence of events taking place during budding and release of Marburg virus. We propose that this represents a general sequence of events also followed by other filamentous and rod-shaped viruses.

  11. Optical properties of bud scales and protochlorophyll(ide) forms in leaf primordia of closed and opened buds.

    Science.gov (United States)

    Solymosi, Katalin; Böddi, Béla

    2006-08-01

    The transmission spectra of bud scales of 14 woody species and the 77 K fluorescence emission spectra of the innermost leaf primordia of closed and opened buds of 37 woody species were studied. Pigment concentrations were determined in some species. Bud scales had low transmittance between 400 and 680 nm with a local minimum around 680 nm. Transmittance increased steeply above 680 nm and was > 80% in the 700-800 nm spectral region. Significant protochlorophyllide (Pchlide) accumulation was observed in leaf primordia of tightly packed, closed buds with relatively thick, dark bud scales. In common ash (Fraxinus excelsior L.) and Hungarian ash (Fraxinus angustifolia Vahl.), the innermost leaf primordia of the closed buds contained protochlorophyll (Pchl) and Pchlide (abbreviated as Pchl(ide)), but no chlorophyll. We observed Pchl(ide) forms with emission maxima at 633, 643 and 655 nm in these leaves. Complete transformation of Pchlide(655) (protochlorophyllide form with maximum emission at 655 nm) into Chlide(692) (chlorophyllide form with maximum emission at 692 nm) occurred after irradiation for 10 s. The innermost leaf primordia of the buds of four species (flowering ash (Fraxinus ornus L.), horse chestnut (Aesculus hippocastanum L.), tree of heaven (Ailanthus altissima P. Mill.) and common walnut (Juglans regia L.)) contained Pchl(ide)(633), Pchl(ide)(643) and Pchlide(655) as well as an emission band at 688 nm corresponding to a chlorophyll form. The Pchlide(655) was fully photoactive in these species. The outermost leaf primordia of these four species and the innermost leaf primordia of 28 other species contained all of the above described Pchl(ide) forms in various ratios but in small amounts. In addition, Chl forms were present and the main bands in the fluorescence emission spectra were at 690 or 740 nm, or both. The results indicate that Pchl(ide) accumulation occurs in leaf primordia in near darkness inside the tightly closed buds, where the bud scales and

  12. Budding yeast colony growth study based on circular granular cell

    Science.gov (United States)

    Aprianti, Devi; Khotimah, S. N.; Viridi, S.

    2016-08-01

    Yeast colony growth can be modelled by using circular granular cells, which can grow and produce buds. The bud growth angle can be set to regulate cell budding pattern. Cohesion force, contact force and Stokes force were adopted to accommodate the behaviour and interactions among cells. Simulation steps are divided into two steps, the explicit step is due to cell growing and implicit step for the cell rearrangement. Only in explicit step that time change was performed. In this study, we examine the influence of cell diameter growth time and reproduction time combination toward the growth of cell number and colony formation. We find a commutative relation between the cell diameter growth time and reproduction time to the specific growth rate. The greater value of the multiplication of the parameters, the smaller specific growth rate is obtained. It also shows a linear correlation between the specific growth rate and colony diameter growth rate.

  13. Adventitious bud regeneration from the stigma of Sinapis alba L.

    Directory of Open Access Journals (Sweden)

    Elżbieta Zenkteler

    2012-12-01

    Full Text Available Stigmas isolated from flower buds of 'Nakielska' variety of Sinapis alba were used to develop a micropropagation method suitable for breeding of new cultivars. The origin of adventitious bud regeneration was studied on MS medium, under stimulation by bezylaminopurine (BAP in combination with 2,4-D - dichlorophenoxyacetic acid (2,4-D. Histological analysis showed the structure of Sinapis stigma (composed from four types of tissue: papillae, transmitting tissue, parenchyma and vascular bundles and revealed that numerous meristematic centers developed from parenchyma cells in close vicinity of vascular bundles. Buds very quickly appeared on the surface of initial explants and later formed multiplantlets that were easily rooted in the soil.

  14. Respiratory Response of Dormant Nectarine Floral Buds on Chilling Deficiency

    Institute of Scientific and Technical Information of China (English)

    TAN Yue; GAO Dong-sheng; LI Ling; CHEN Xiu-de; XU Ai-hong

    2010-01-01

    Changes in main biochemical respiratory pathways in dormant nectarine floral buds were studied with nectarine trees (Prunus persica.var,nectariana cv.Shuguang) in order to determine the function of respiration in dormancy release.Oxygen-electrode system and respiratory inhibitors were used to measure total respiratory rates and rates of respiratory pathways.Results showed that chilling deficiency blocked the transition of respiratory mode,and made buds stay in a state of high level pentose phosphate pathway (PPP) and low level tricarboxylic acid cycle (TCA).The decline of PPP and activation of TCA occurred synchronously with the release of dormancy.In addition,the inhibition of PPP stimulated a respiration increase related with TCA.It could be concluded that the function of PPP activation in dormancy release might be limited and PPP declination inducing TCA activation might be part of respiration mode transition mechanism during bud sprouting.

  15. GABA, its receptors, and GABAergic inhibition in mouse taste buds

    OpenAIRE

    Dvoryanchikov, Gennady; Huang, Yijen A.; Barro-Soria, Rene; Chaudhari, Nirupa; Roper, Stephen D.

    2011-01-01

    Taste buds consist of at least three principal cell types that have different functions in processing gustatory signals — glial-like Type I cells, Receptor (Type II) cells, and Presynaptic (Type III) cells. Using a combination of Ca2+ imaging, single cell RT-PCR, and immunostaining, we show that γ-amino butyric acid (GABA) is an inhibitory transmitter in mouse taste buds, acting on GABA-A and GABA-B receptors to suppress transmitter (ATP) secretion from Receptor cells during taste stimulation...

  16. RESEARCH OF SOPHORA JAPONICA L. FLOWER BUDS VOLATILE COMPOUNDS WITH GAS-CHROMATOGRAPHY/MASS- SPECTROMETRY METHOD

    Directory of Open Access Journals (Sweden)

    Cholak I.S.

    2013-10-01

    Full Text Available This work represents the results of the research ofessential oil contained in Sophora japonica L. flowerbuds volatile compounds collected during the nextstages of their development: green flower buds, formedflower buds and the beginning of flower buds opening.Essential oil assay content in Sophora japonica L.flower buds was determined with hydrodistillationmethod. Content of essential oil in the raw material isless than 0,1%. Qualitative composition and assaycontent of Sophora japonica L. flower buds essential oilconstituents were determined with chromato-massspectrometry method. In consequence of the research 80constituents were identified in Sophora japonica L.flower buds out of which 61 substances are during thegreen flower buds and beginning of flower budsopening stages, 66 substances are during formed flowerbuds stage. Substances are represented by aliphatic andcyclic terpenoids, their alcohols and ketones. Mostvolatile substances were extracted on the stage offormed buds.

  17. Effect of gamma irradiated parenchyma on the growth of irradiated potato tuber buds

    International Nuclear Information System (INIS)

    The development of buds greffed on irradiated potato parenchyma was studied. The irradiated parenchyma does not influence the sprouting capacity of buds, but it affects the way they develop. (Author) 9 refs

  18. A New Compound from the Bud of Chrysanthemum indicum L.

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    A new bicyclic spiroketone was isolated from the bud of Chrysanthemum indicum L.The chemical structure was elucidated as (1R, 9S, 10S)-10-hydroxyl-8 (2', 4'-diynehexylidene)-9-isovaleryloxy-2, 7-dioxaspiro [5, 4] decane based on the X-ray crystallography.

  19. Axillary Bud Proliferation Approach for Plant Biodiversity Conservation and Restoration

    Directory of Open Access Journals (Sweden)

    F. Ngezahayo

    2014-01-01

    Full Text Available Due to mainly human population pressure and activities, global biodiversity is getting reduced and particularly plant biodiversity is becoming at high risk of extinction. Consequently, many efforts have been deployed to develop conservation methods. Because it does not involve cell dedifferentiation of differentiated cells but rather the development and growth of new shoots from preexisting meristems, the axillary bud proliferation approach is the method offering least risk of genetic instability. Indeed, meristems are more resistant to genetic changes than disorganized tissues. The present review explored through the scientific literature the axillary bud proliferation approach and the possible somaclonal variation that could arise from it. Almost genetic stability or low level of genetic variation is often reported. On the contrary, in a few cases studied to date, DNA methylation alterations often appeared in the progenies, showing epigenetic variations in the regenerated plants from axillary bud culture. Fortunately, epigenetic changes are often temporary and plants may revert to the normal phenotype. Thus, in the absence of genetic variations and the existence of reverting epigenetic changes over time, axillary bud culture can be adopted as an alternative nonconventional way of conserving and restoring of plant biodiversity.

  20. Regulation of homologous recombination at telomeres in budding yeast

    DEFF Research Database (Denmark)

    Eckert-Boulet, Nadine; Lisby, Michael

    2010-01-01

    Homologous recombination is suppressed at normal length telomere sequences. In contrast, telomere recombination is allowed when telomeres erode in the absence of telomerase activity or as a consequence of nucleolytic degradation or incomplete replication. Here, we review the mechanisms...... that contribute to regulating mitotic homologous recombination at telomeres and the role of these mechanisms in signalling short telomeres in the budding yeast Saccharomyces cerevisiae....

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    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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  9. Induction of ectopic taste buds by SHH reveals the competency and plasticity of adult lingual epithelium.

    Science.gov (United States)

    Castillo, David; Seidel, Kerstin; Salcedo, Ernesto; Ahn, Christina; de Sauvage, Frederic J; Klein, Ophir D; Barlow, Linda A

    2014-08-01

    Taste buds are assemblies of elongated epithelial cells, which are innervated by gustatory nerves that transmit taste information to the brain stem. Taste cells are continuously renewed throughout life via proliferation of epithelial progenitors, but the molecular regulation of this process remains unknown. During embryogenesis, sonic hedgehog (SHH) negatively regulates taste bud patterning, such that inhibition of SHH causes the formation of more and larger taste bud primordia, including in regions of the tongue normally devoid of taste buds. Here, using a Cre-lox system to drive constitutive expression of SHH, we identify the effects of SHH on the lingual epithelium of adult mice. We show that misexpression of SHH transforms lingual epithelial cell fate, such that daughter cells of lingual epithelial progenitors form cell type-replete, onion-shaped taste buds, rather than non-taste, pseudostratified epithelium. These SHH-induced ectopic taste buds are found in regions of the adult tongue previously thought incapable of generating taste organs. The ectopic buds are composed of all taste cell types, including support cells and detectors of sweet, bitter, umami, salt and sour, and recapitulate the molecular differentiation process of endogenous taste buds. In contrast to the well-established nerve dependence of endogenous taste buds, however, ectopic taste buds form independently of both gustatory and somatosensory innervation. As innervation is required for SHH expression by endogenous taste buds, our data suggest that SHH can replace the need for innervation to drive the entire program of taste bud differentiation. PMID:24993944

  10. Membrane-elasticity model of Coatless vesicle budding induced by ESCRT complexes.

    Directory of Open Access Journals (Sweden)

    Bartosz Różycki

    Full Text Available The formation of vesicles is essential for many biological processes, in particular for the trafficking of membrane proteins within cells. The Endosomal Sorting Complex Required for Transport (ESCRT directs membrane budding away from the cytosol. Unlike other vesicle formation pathways, the ESCRT-mediated budding occurs without a protein coat. Here, we propose a minimal model of ESCRT-induced vesicle budding. Our model is based on recent experimental observations from direct fluorescence microscopy imaging that show ESCRT proteins colocalized only in the neck region of membrane buds. The model, cast in the framework of membrane elasticity theory, reproduces the experimentally observed vesicle morphologies with physically meaningful parameters. In this parameter range, the minimum energy configurations of the membrane are coatless buds with ESCRTs localized in the bud neck, consistent with experiment. The minimum energy configurations agree with those seen in the fluorescence images, with respect to both bud shapes and ESCRT protein localization. On the basis of our model, we identify distinct mechanistic pathways for the ESCRT-mediated budding process. The bud size is determined by membrane material parameters, explaining the narrow yet different bud size distributions in vitro and in vivo. Our membrane elasticity model thus sheds light on the energetics and possible mechanisms of ESCRT-induced membrane budding.

  11. Influences of polar auxin transport on polarity of adventitious bud formation in hybrid populas

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Myung Won (Yonsei Univ. Kangwondo (Korea)); Hackett, W. (Univ of Minnesota, St. Paul (USA))

    1989-04-01

    The role of auxin and cytokinin distribution of polar regeneration of adventitious bud has been investigated. Explants from leaf midvein were labelled with {sup 14}C-NAA and {sup 14}C-BA and the radioactivity in proximal, mid, and distal portions was counted after 24h and 48h. Explants showing polar regeneration of buds on the proximal end showed a clear polar distribution of {sup 14}CNAA. Auxin transport inhibitors (NPA, TIBA) eliminated polar distribution of auxin and reduced polarity of bud formation and the total number of buds formed, but did not reduce callus formation. Increased concentration of Ca(NO{sub 3}){sub 2} decreased polarity of bud formation and increased the number of buds formed but did not affect the distribution of auxin of cytokinin. Some factor in addition to polar distribution of auxin or cytokinin-auxin ratio appears to influence the polarity of adventitious bud formation.

  12. Tumor budding is a strong and reproducible prognostic marker in T3N0 colorectal cancer.

    LENUS (Irish Health Repository)

    Wang, Lai Mun

    2012-02-01

    BACKGROUND: Tumor budding along the advancing front of colorectal adenocarcinoma is an early event in the metastatic process. A reproducible, prognostic budding scoring system based on outcomes in early stage colorectal cancer has not been established. DESIGN: One hundred twenty-eight T3N0M0 colorectal carcinoma patients with known outcome were identified. Tumor budding was defined as isolated tumor cells or clusters of <5 cells at the invasive tumor front. Tumor bud counts were generated in 5 regions at 200x by 2 pathologists (conventional bud count method). The median bud count per case was used to divide cases into low (median=0) and high budding (median > or =1) groups. Forty cases were reevaluated to assess reproducibility using the conventional and a novel rapid bud count method. RESULTS: Fifty-seven (45%) carcinomas had high and 71 (55%) had low budding scores. High budding was associated with an infiltrative growth pattern (P<0.0001) and lymphovascular invasion (P=0.005). Five-year cancer-specific survival was significantly poorer in high compared with low budding groups: 63% versus 91%, respectively, P<0.0001. Multivariate analysis demonstrated tumor budding to be independently prognostic (hazard ratio=4.76, P<0.001). Interobserver agreement was at least equivalent comparing the conventional to the rapid bud count methods: 87.5% agreement (kappa=0.75) versus 92.5% agreement (kappa=0.85), respectively. CONCLUSIONS: Tumor budding is a strong, reproducible, and independent prognostic marker of outcome that is easily assessed on hematoxylin and eosin slides. This may be useful for identifying the subset of T3N0M0 patients at high risk of recurrence who may benefit from adjuvant therapy.

  13. Recurrent peripheral odontogenic fibroma associated with basal cell budding.

    Science.gov (United States)

    Sreeja, C; Vezhavendan, N; Shabana, F; Vijayalakshmi, D; Devi, M; Arunakiry, N

    2014-07-01

    Peripheral odontogenic fibroma (POdF) is a rare benign odontogenic neoplasm. It represents the soft tissue counterpart of central odontogenic fibroma. The embryonic source of POdF has been suggested by many as arising from the rest of dental lamina that has persisted in the gingiva following its disintegration. It presents clinically as a firm, slow growing and sessile gingival mass, which is difficult to distinguish with more common inflammatory lesions. Very few cases of recurrence have been documented. It has been stated that histological budding of basal cell layer of the surface squamous epithelium is associated with higher recurrence and the presence of calcification in direct apposition to the epithelial rest is associated with lower recurrence. Hereby, we present a case which histologically exhibited budding of the basal cell layer, which could have been the reason for its recurrence. PMID:25210375

  14. Recurrent peripheral odontogenic fibroma associated with basal cell budding

    Directory of Open Access Journals (Sweden)

    C Sreeja

    2014-01-01

    Full Text Available Peripheral odontogenic fibroma (POdF is a rare benign odontogenic neoplasm. It represents the soft tissue counterpart of central odontogenic fibroma. The embryonic source of POdF has been suggested by many as arising from the rest of dental lamina that has persisted in the gingiva following its disintegration. It presents clinically as a firm, slow growing and sessile gingival mass, which is difficult to distinguish with more common inflammatory lesions. Very few cases of recurrence have been documented. It has been stated that histological budding of basal cell layer of the surface squamous epithelium is associated with higher recurrence and the presence of calcification in direct apposition to the epithelial rest is associated with lower recurrence. Hereby, we present a case which histologically exhibited budding of the basal cell layer, which could have been the reason for its recurrence.

  15. Budding Transition of Asymmetric Two-component Lipid Domains

    CERN Document Server

    Wolff, Jean; Andelman, David

    2016-01-01

    We propose a model that accounts for the budding transition of asymmetric two-component lipid domains, where the two monolayers (leaflets) have different average compositions controlled by independent chemical potentials. Assuming a coupling between the local curvature and local lipid composition in each of the leaflets, we discuss the morphology and thermodynamic behavior of asymmetric lipid domains. The membrane free-energy contains three contributions: the bending energy, the line tension, and a Landau free-energy for a lateral phase separation. Within a mean-field treatment, we obtain various phase diagrams containing fully budded, dimpled, and flat states as a function of the two leaflet compositions. The global phase behavior is analyzed, and depending on system parameters, the phase diagrams include one-phase, two-phase and three-phase regions. In particular, we predict various phase coexistence regions between different morphologies of domains, which may be observed in multi-component membranes or ves...

  16. Newly identified prions in budding yeast, and their possible functions

    OpenAIRE

    Crow, Emily T.; Li, Liming

    2011-01-01

    Yeast prions are atypical genetic elements that are transmitted as heritable protein conformations. [PSI+], [URE3], and [PIN+] are three well-studied prions in the budding yeast, Saccharomyces cerevisiae. In the last three years, several additional prions have been reported in yeast, including [SWI+], [OCT+], [MCA], [GAR+], [MOT3+], [ISP+], and [NSI+]. The growing number of yeast prions suggests that protein-based inheritance might be a widespread biological phenomenon. In this review, we sum...

  17. Endogenous peripheral neuromodulators of the mammalian taste bud.

    Science.gov (United States)

    Dando, Robin

    2010-10-01

    The sensitivity of the mammalian taste system displays a degree of plasticity based on short-term nutritional requirements. Deficiency in a particular substance may lead to a perceived increase in palatability of this substance, providing an additional drive to redress this nutritional imbalance through modification of intake. This alteration occurs not only in the brain but also, before any higher level processing has occurred, in the taste buds themselves. A brief review of recent advances is offered.

  18. Measuring Replicative Life Span in the Budding Yeast

    OpenAIRE

    Steffen, Kristan K.; Kennedy, Brian K.; Kaeberlein, Matt

    2009-01-01

    Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The budding yeast Saccharomyces cerevisiae has been used extensively to study the biology of aging, and several determinants of yeast longevity have been shown to be conserved in multicellular eukaryotes, including worms, flies, and mice 1. Due to the lack of easily quantified age-associated phenotypes, aging in yeast has been assayed almost exclusively by...

  19. Evaluation and Properties of the Budding Yeast Phosphoproteome

    OpenAIRE

    Amoutzias, G. D.; He, Y.; Lilley, K. S.; Van de Peer, Y.; Oliver, S G

    2012-01-01

    We have assembled a reliable phosphoproteomic data set for budding yeast Saccharomyces cerevisiae and have investigated its properties. Twelve publicly available phosphoproteome data sets were triaged to obtain a subset of high-confidence phosphorylation sites (p-sites), free of "noisy" phosphorylations. Analysis of this combined data set suggests that the inventory of phosphoproteins in yeast is close to completion, but that these proteins may have many undiscovered p-sites. Proteins involve...

  20. The protein machinery of vesicle budding and fusion.

    OpenAIRE

    Rothman, J E

    1996-01-01

    A general protein machinery that buds and fuses transport vesicles is harnessed to generate the complex web of intracellular transport pathways critical for such diverse processes as cell growth, endocytosis, hormone release, and neurotransmission. With this appreciation, the challenge of understanding the precise molecular mechanisms of these many facets of cell biology has been reduced to a series of problems in protein structure and chemistry.

  1. Signal transduction and information processing in mammalian taste buds

    OpenAIRE

    Roper, Stephen D.

    2007-01-01

    The molecular machinery for chemosensory transduction in taste buds has received considerable attention within the last decade. Consequently, we now know a great deal about sweet, bitter, and umami taste mechanisms and are gaining ground rapidly on salty and sour transduction. Sweet, bitter, and umami tastes are transduced by G-protein-coupled receptors. Salty taste may be transduced by epithelial Na channels similar to those found in renal tissues. Sour transduction appears to be initiated b...

  2. Ecological conditions favoring budding in colonial organisms under environmental disturbance.

    Directory of Open Access Journals (Sweden)

    Mayuko Nakamaru

    Full Text Available Dispersal is a topic of great interest in ecology. Many organisms adopt one of two distinct dispersal tactics at reproduction: the production of small offspring that can disperse over long distances (such as seeds and spawned eggs, or budding. The latter is observed in some colonial organisms, such as clonal plants, corals and ants, in which (superorganisms split their body into components of relatively large size that disperse to a short distance. Contrary to the common dispersal viewpoint, short-dispersal colonial organisms often flourish even in environments with frequent disturbances. In this paper, we investigate the conditions that favor budding over long-distance dispersal of small offspring, focusing on the life history of the colony growth and the colony division ratio. These conditions are the relatively high mortality of very small colonies, logistic growth, the ability of dispersers to peacefully seek and settle unoccupied spaces, and small spatial scale of environmental disturbance. If these conditions hold, budding is advantageous even when environmental disturbance is frequent. These results suggest that the demography or life history of the colony underlies the behaviors of the colonial organisms.

  3. Bud dormancy in apple trees after thermal fluctuations

    Directory of Open Access Journals (Sweden)

    Rafael Anzanello

    2014-06-01

    Full Text Available The objective of this work was to evaluate the effect of heat waves on the evolution of bud dormancy, in apple trees with contrasting chilling requirements. Twigs of 'Castel Gala' and 'Royal Gala' were collected in orchards in Papanduva, state of Santa Catarina, Brazil, and were exposed to constant (3°C or alternating (3 and 15°C for 12/12 hours temperature, combined with zero, one or two days a week at 25°C. Two additional treatments were evaluated: constant temperature (3°C, with a heat wave of seven days at 25°C, in the beginning or in the middle of the experimental period. Periodically, part of the twigs was transferred to 25°C for daily budburst evaluation of apical and lateral buds. Endodormancy (dormancy induced by cold was overcome with less than 330 chilling hours (CH of constant cold in 'Castel Gala' and less than 618 CH in 'Royal Gala'. A daily 15°C-temperature cycle did not affect the endodormancy process. Heat waves during endodormancy resulted in an increased CH to achieve bud requirements. The negative effect of high temperature depended on the lasting of this condition. Chilling was partly cancelled during dormancy when the heat wave lasted 36 continuous hours or more. Therefore, budburst prediction models need adjustments, mainly for regions with mild and irregular winters, such as those of Southern Brazil.

  4. Trichomes control flower bud shape by linking together young petals.

    Science.gov (United States)

    Tan, Jiafu; Walford, Sally-Anne; Dennis, Elizabeth S; Llewellyn, Danny

    2016-01-01

    Trichomes are widespread in plants and develop from surface cells on different tissues(1). They have many forms and functions, from defensive spines to physical barriers that trap layers of air to insulate against desiccation, but there is growing evidence that trichomes can also have developmental roles in regulating flower structure(2,3). We report here that the trichomes on petals of cotton, Gossypium hirsutum L., are essential for correct flower bud shape through a mechanical entanglement of the trichomes on adjacent petals that anchor the edges to counter the opposing force generated by asymmetric expansion of overlapping petals. Silencing a master regulator of petal trichomes, GhMYB-MIXTA-Like10 (GhMYBML10), by RNA interference (RNAi) suppressed petal trichome growth and resulted in flower buds forming into abnormal corkscrew shapes that exposed developing anthers and stigmas to desiccation damage. Artificially gluing petal edges together could partially restore correct bud shape and fertility. Such petal 'Velcro' is present in other Malvaceae and perhaps more broadly in other plant families, although it is not ubiquitous. This mechanism for physical association between separate organs to regulate flower shape and function is different from the usual organ shape control(4) exerted through cell-to-cell communication and differential cell expansion within floral tissues(5,6). PMID:27322517

  5. Strigolactone acts downstream of auxin to regulate bud outgrowth in pea and Arabidopsis.

    Science.gov (United States)

    Brewer, Philip B; Dun, Elizabeth A; Ferguson, Brett J; Rameau, Catherine; Beveridge, Christine A

    2009-05-01

    During the last century, two key hypotheses have been proposed to explain apical dominance in plants: auxin promotes the production of a second messenger that moves up into buds to repress their outgrowth, and auxin saturation in the stem inhibits auxin transport from buds, thereby inhibiting bud outgrowth. The recent discovery of strigolactone as the novel shoot-branching inhibitor allowed us to test its mode of action in relation to these hypotheses. We found that exogenously applied strigolactone inhibited bud outgrowth in pea (Pisum sativum) even when auxin was depleted after decapitation. We also found that strigolactone application reduced branching in Arabidopsis (Arabidopsis thaliana) auxin response mutants, suggesting that auxin may act through strigolactones to facilitate apical dominance. Moreover, strigolactone application to tiny buds of mutant or decapitated pea plants rapidly stopped outgrowth, in contrast to applying N-1-naphthylphthalamic acid (NPA), an auxin transport inhibitor, which significantly slowed growth only after several days. Whereas strigolactone or NPA applied to growing buds reduced bud length, only NPA blocked auxin transport in the bud. Wild-type and strigolactone biosynthesis mutant pea and Arabidopsis shoots were capable of instantly transporting additional amounts of auxin in excess of endogenous levels, contrary to predictions of auxin transport models. These data suggest that strigolactone does not act primarily by affecting auxin transport from buds. Rather, the primary repressor of bud outgrowth appears to be the auxin-dependent production of strigolactones. PMID:19321710

  6. Temperature Characteristics of Dormancy Development of Terminal and Lateral Buds in Paulownia

    Institute of Scientific and Technical Information of China (English)

    LIUZhen; LIGuangtao; WANGYanmei; JIANGXiaoping

    2004-01-01

    In order to probe the death reason of terminal buds and carry out trunk extension by using lateral buds, the temperature characteristics of dormancy development of terminal and lateral buds in Paulownia tomentosa xp. fortunei 33 were investigated by raising cutting branches on different date at 25℃ and 15℃. The results were as follows:①The death reason of terminal buds and the first pair of lateral buds was itselfe cological adapting strategy, not the injury of early frost and freezing; ② The lateral buds 2, 3 and 4 had obtained the feature of winter dormancy, and could survive from cold winter, and widen its possible temperature range of sprouting to lower temperature by experiencing winter chilling; ③The lateral buds 2, 3 and 4 could sprout at the same time in spring, but it was difficult to sprout for other lateral buds; ④ The sprouting of upper lateral buds could restrict the sprouting of lower lateral buds at 25℃, but not at 15℃.

  7. Differentiation of Apical Bud Cells in a Newly Developed Apical Bud Transplantation Model Using GFP Transgenic Mice as Donor

    Science.gov (United States)

    Sakagami, Ryuji; Yoshinaga, Yasunori; Okamura, Kazuhiko

    2016-01-01

    Rodent mandibular incisors have a unique anatomical structure that allows teeth to grow throughout the lifetime of the rodent. This report presents a novel transplantation technique for studying the apical bud differentiation of rodent mandibular incisors. Incisal apical end tissue with green fluorescent protein from transgenic mouse was transplanted to wild type mice, and the development of the transplanted cells were immunohistologically observed for 12 weeks after the transplantation. Results indicate that the green fluorescent apical end tissue replaced the original tissue, and cells from the apical bud differentiated and extended toward the incisal edge direction. The immunostaining with podoplanin also showed that the characteristics of the green fluorescent tissue were identical to those of the original. The green fluorescent cells were only found in the labial side of the incisor up to 4 weeks. After 12 weeks, however, they were also found in the lingual side. Here the green fluorescent cementocyte-like cells were only present in the cementum close to the dentin surface. This study suggests that some of the cells that form the cellular cementum come from the apical tissue including the apical bud in rodent incisors. PMID:26978064

  8. Genome-wide analysis of gene expression in primate taste buds reveals links to diverse processes.

    Directory of Open Access Journals (Sweden)

    Peter Hevezi

    Full Text Available Efforts to unravel the mechanisms underlying taste sensation (gustation have largely focused on rodents. Here we present the first comprehensive characterization of gene expression in primate taste buds. Our findings reveal unique new insights into the biology of taste buds. We generated a taste bud gene expression database using laser capture microdissection (LCM procured fungiform (FG and circumvallate (CV taste buds from primates. We also used LCM to collect the top and bottom portions of CV taste buds. Affymetrix genome wide arrays were used to analyze gene expression in all samples. Known taste receptors are preferentially expressed in the top portion of taste buds. Genes associated with the cell cycle and stem cells are preferentially expressed in the bottom portion of taste buds, suggesting that precursor cells are located there. Several chemokines including CXCL14 and CXCL8 are among the highest expressed genes in taste buds, indicating that immune system related processes are active in taste buds. Several genes expressed specifically in endocrine glands including growth hormone releasing hormone and its receptor are also strongly expressed in taste buds, suggesting a link between metabolism and taste. Cell type-specific expression of transcription factors and signaling molecules involved in cell fate, including KIT, reveals the taste bud as an active site of cell regeneration, differentiation, and development. IKBKAP, a gene mutated in familial dysautonomia, a disease that results in loss of taste buds, is expressed in taste cells that communicate with afferent nerve fibers via synaptic transmission. This database highlights the power of LCM coupled with transcriptional profiling to dissect the molecular composition of normal tissues, represents the most comprehensive molecular analysis of primate taste buds to date, and provides a foundation for further studies in diverse aspects of taste biology.

  9. Ice nucleation activity in various tissues of Rhododendron flower buds: their relevance to extraorgan freezing

    Directory of Open Access Journals (Sweden)

    Masaya eIshikawa

    2015-03-01

    Full Text Available Wintering flower buds of cold hardy Rhododendron japonicum cooled slowly to subfreezing temperatures are known to undergo extraorgan freezing, whose mechanisms remain obscure. We revisited this material to demonstrate why bud scales freeze first in spite of their lower water content, why florets remain deeply supercooled and how seasonal adaptive responses occur in regard to extraorgan freezing in flower buds. We determined ice nucleation activity (INA of various flower bud tissues of using a test tube-based assay. Irrespective of collection sites, outer and inner bud scales that function as ice sinks in extraorgan freezing had high INA levels whilst florets that remain supercooled and act as a water source lacked INA. The INA level of bud scales was not high in late August when flower bud formation was ending, but increased to reach the highest level in late October just before the first autumnal freeze. The results support the following hypothesis: the high INA in bud scales functions as the subfreezing sensor, ensuring the primary freezing in bud scales at warmer subzero temperatures, which likely allows the migration of floret water to the bud scales and accumulation of icicles within the bud scales. The low INA in the florets helps them remain unfrozen by deep supercooling. The INA in the bud scales was resistant to grinding and autoclaving at 121°C for 15 min, implying the intrinsic nature of the INA rather than of microbial origin, whilst the INA in stem bark was autoclaving labile. Anti-nucleation activity (ANA was implicated in the leachate of autoclaved bud scales, which suppresses the INA at millimolar levels of concentration and likely differs from the colligative effects of the solutes. The tissue INA levels likely contribute to the establishment of freezing behaviors by ensuring the order of freezing in the tissues: from the primary freeze to the last tissue remaining unfrozen.

  10. Ice nucleation activity in various tissues of Rhododendron flower buds: their relevance to extraorgan freezing.

    Science.gov (United States)

    Ishikawa, Masaya; Ishikawa, Mikiko; Toyomasu, Takayuki; Aoki, Takayuki; Price, William S

    2015-01-01

    Wintering flower buds of cold hardy Rhododendron japonicum cooled slowly to subfreezing temperatures are known to undergo extraorgan freezing, whose mechanisms remain obscure. We revisited this material to demonstrate why bud scales freeze first in spite of their lower water content, why florets remain deeply supercooled and how seasonal adaptive responses occur in regard to extraorgan freezing in flower buds. We determined ice nucleation activity (INA) of various flower bud tissues using a test tube-based assay. Irrespective of collection sites, outer and inner bud scales that function as ice sinks in extraorgan freezing had high INA levels whilst florets that remain supercooled and act as a water source lacked INA. The INA level of bud scales was not high in late August when flower bud formation was ending, but increased to reach the highest level in late October just before the first autumnal freeze. The results support the following hypothesis: the high INA in bud scales functions as the subfreezing sensor, ensuring the primary freezing in bud scales at warmer subzero temperatures, which likely allows the migration of floret water to the bud scales and accumulation of icicles within the bud scales. The low INA in the florets helps them remain unfrozen by deep supercooling. The INA in the bud scales was resistant to grinding and autoclaving at 121(∘)C for 15 min, implying the intrinsic nature of the INA rather than of microbial origin, whilst the INA in stem bark was autoclaving-labile. Anti-nucleation activity (ANA) was implicated in the leachate of autoclaved bud scales, which suppresses the INA at millimolar levels of concentration and likely differs from the colligative effects of the solutes. The tissue INA levels likely contribute to the establishment of freezing behaviors by ensuring the order of freezing in the tissues: from the primary freeze to the last tissue remaining unfrozen. PMID:25859249

  11. Involvement of EARLY BUD-BREAK, an AP2/ERF Transcription Factor Gene, in Bud Break in Japanese Pear (Pyrus pyrifolia Nakai) Lateral Flower Buds: Expression, Histone Modifications and Possible Target Genes.

    Science.gov (United States)

    Anh Tuan, Pham; Bai, Songling; Saito, Takanori; Imai, Tsuyoshi; Ito, Akiko; Moriguchi, Takaya

    2016-05-01

    In the Japanese pear (Pyrus pyrifolia Nakai) 'Kosui', three developmental stages of lateral flower buds have been proposed to occur during ecodormancy to the flowering phase, i.e. rapid enlargement, sprouting and flowering. Here, we report an APETALA2/ethylene-responsive factor (AP2/ERF) transcription factor gene, named pear EARLY BUD-BREAK (PpEBB), which was highly expressed during the rapid enlargement stage occurring prior to the onset of bud break in flower buds. Gene expression analysis revealed that PpEBB expression was dramatically increased during the rapid enlargement stage in three successive growing seasons. PpEBB transcript levels peaked 1 week prior to onset of bud break in 'Kosui' potted plants treated with hydrogen cyanamide or water under forcing conditions. Chromatin immunoprecipitation-quantitative PCR showed that higher levels of active histone modifications (trimethylation of the histone H3 tail at Lys4) in the 5'-upstream and start codon regions of the PpEBB gene were associated with the induced expression level of PpEBB during the rapid enlargement stage. In addition, we provide evidence that PpEBB may interact with and regulate pear four D-type cyclin (PpCYCD3) genes during bud break in 'Kosui' lateral flower buds. PpEBB significantly increased the promoter activities of four PpCYCD3 genes in a dual-luciferase assay using tobacco leaves. Taken together, our findings uncovered aspects of the bud break regulatory mechanism in the Japanese pear and provided further evidence that the EBB family plays an important role in bud break in perennial plants. PMID:26940832

  12. Reconstituting ring-rafts in bud-mimicking topography of model membranes

    Science.gov (United States)

    Ryu, Yong-Sang; Lee, In-Ho; Suh, Jeng-Hun; Park, Seung Chul; Oh, Soojung; Jordan, Luke R.; Wittenberg, Nathan J.; Oh, Sang-Hyun; Jeon, Noo Li; Lee, Byoungho; Parikh, Atul N.; Lee, Sin-Doo

    2014-07-01

    During vesicular trafficking and release of enveloped viruses, the budding and fission processes dynamically remodel the donor cell membrane in a protein- or a lipid-mediated manner. In all cases, in addition to the generation or relief of the curvature stress, the buds recruit specific lipids and proteins from the donor membrane through restricted diffusion for the development of a ring-type raft domain of closed topology. Here, by reconstituting the bud topography in a model membrane, we demonstrate the preferential localization of cholesterol- and sphingomyelin-enriched microdomains in the collar band of the bud-neck interfaced with the donor membrane. The geometrical approach to the recapitulation of the dynamic membrane reorganization, resulting from the local radii of curvatures from nanometre-to-micrometre scales, offers important clues for understanding the active roles of the bud topography in the sorting and migration machinery of key signalling proteins involved in membrane budding.

  13. Membrane tension is a key determinant of bud morphology in clathrin-mediated endocytosis

    CERN Document Server

    Hassinger, Julian E; Drubin, David G; Rangamani, Padmini

    2016-01-01

    In clathrin-mediated endocytosis (CME), clathrin and various adaptor proteins coat a patch of the plasma membrane, which is reshaped to form a budded vesicle. Experimental studies have demonstrated that elevated membrane tension can inhibit bud formation by a clathrin coat. In this study, we investigate the impact of membrane tension on the mechanics of membrane budding by simulating clathrin coats that either grow in area or progressively induce greater curvature. At low membrane tension, progressively increasing the area of a curvature-generating coat causes the membrane to smoothly evolve from a flat to budded morphology, whereas the membrane remains essentially flat at high membrane tensions. Interestingly, at physiologically relevant, intermediate membrane tensions, the shape evolution of the membrane undergoes a snapthrough instability in which increasing coat area causes the membrane to "snap" from an open, U-shaped bud to a closed, $\\Omega$-shaped bud. This instability is accompanied by a large energy...

  14. Sucrose is an early modulator of the key hormonal mechanisms controlling bud outgrowth in Rosa hybrida.

    Science.gov (United States)

    Barbier, François; Péron, Thomas; Lecerf, Marion; Perez-Garcia, Maria-Dolores; Barrière, Quentin; Rolčík, Jakub; Boutet-Mercey, Stéphanie; Citerne, Sylvie; Lemoine, Remi; Porcheron, Benoît; Roman, Hanaé; Leduc, Nathalie; Le Gourrierec, José; Bertheloot, Jessica; Sakr, Soulaiman

    2015-05-01

    Sugar has only recently been identified as a key player in triggering bud outgrowth, while hormonal control of bud outgrowth is already well established. To get a better understanding of sugar control, the present study investigated how sugar availability modulates the hormonal network during bud outgrowth in Rosa hybrida. Other plant models, for which mutants are available, were used when necessary. Buds were grown in vitro to manipulate available sugars. The temporal patterns of the hormonal regulatory network were assessed in parallel with bud outgrowth dynamics. Sucrose determined bud entrance into sustained growth in a concentration-dependent manner. Sustained growth was accompanied by sustained auxin production in buds, and sustained auxin export in a DR5::GUS-expressing pea line. Several events occurred ahead of sucrose-stimulated bud outgrowth. Sucrose upregulated early auxin synthesis genes (RhTAR1, RhYUC1) and the auxin efflux carrier gene RhPIN1, and promoted PIN1 abundance at the plasma membrane in a pPIN1::PIN1-GFP-expressing tomato line. Sucrose downregulated both RwMAX2, involved in the strigolactone-transduction pathway, and RhBRC1, a repressor of branching, at an early stage. The presence of sucrose also increased stem cytokinin content, but sucrose-promoted bud outgrowth was not related to that pathway. In these processes, several non-metabolizable sucrose analogues induced sustained bud outgrowth in R. hybrida, Pisum sativum, and Arabidopsis thaliana, suggesting that sucrose was involved in a signalling pathway. In conclusion, we identified potential hormonal candidates for bud outgrowth control by sugar. They are central to future investigations aimed at disentangling the processes that underlie regulation of bud outgrowth by sugar.

  15. Breast bud detection: a validation study in the Chilean Growth Obesity Cohort Study

    OpenAIRE

    Pereira, Ana; Garmendia, María Luisa; González, Daniela; Kain, Juliana; Mericq, Verónica; Uauy, Ricardo; Corvalán, Camila

    2014-01-01

    Background Early puberty onset has been related to future chronic disease; however breast bud assessment in large scale population studies is difficult because it requires trained personnel. Thus our aim is to assess the validity of self and maternal breast bud detection, considering girl’s body mass index (BMI) and maternal education. Methods In 2010, 481 girls (mean age = 7.8) from the Growth and Obesity Chilean Cohort Study were evaluated by a nutritionist trained in breast bud detection. ...

  16. Relationship between formation of gametophore buds in the protonema of mosses and increase in ribonuclease activity

    Directory of Open Access Journals (Sweden)

    M. Spychała

    2015-05-01

    Full Text Available Changes in RNase activity similar to those accompanying cytokinin-induced formation of gametophore buds in mosses (a decrease in the early phase of bud formation and later an increase in enzyme activity have also been found during spontaneous formation of gametophores in moss ontogenesis. Using various factors affecting the cytokinin-induced process of bud formation a correlation has been found between this process and the increase in RNase activity.

  17. Sucrose is an early modulator of the key hormonal mechanisms controlling bud outgrowth in Rosa hybrida.

    Science.gov (United States)

    Barbier, François; Péron, Thomas; Lecerf, Marion; Perez-Garcia, Maria-Dolores; Barrière, Quentin; Rolčík, Jakub; Boutet-Mercey, Stéphanie; Citerne, Sylvie; Lemoine, Remi; Porcheron, Benoît; Roman, Hanaé; Leduc, Nathalie; Le Gourrierec, José; Bertheloot, Jessica; Sakr, Soulaiman

    2015-05-01

    Sugar has only recently been identified as a key player in triggering bud outgrowth, while hormonal control of bud outgrowth is already well established. To get a better understanding of sugar control, the present study investigated how sugar availability modulates the hormonal network during bud outgrowth in Rosa hybrida. Other plant models, for which mutants are available, were used when necessary. Buds were grown in vitro to manipulate available sugars. The temporal patterns of the hormonal regulatory network were assessed in parallel with bud outgrowth dynamics. Sucrose determined bud entrance into sustained growth in a concentration-dependent manner. Sustained growth was accompanied by sustained auxin production in buds, and sustained auxin export in a DR5::GUS-expressing pea line. Several events occurred ahead of sucrose-stimulated bud outgrowth. Sucrose upregulated early auxin synthesis genes (RhTAR1, RhYUC1) and the auxin efflux carrier gene RhPIN1, and promoted PIN1 abundance at the plasma membrane in a pPIN1::PIN1-GFP-expressing tomato line. Sucrose downregulated both RwMAX2, involved in the strigolactone-transduction pathway, and RhBRC1, a repressor of branching, at an early stage. The presence of sucrose also increased stem cytokinin content, but sucrose-promoted bud outgrowth was not related to that pathway. In these processes, several non-metabolizable sucrose analogues induced sustained bud outgrowth in R. hybrida, Pisum sativum, and Arabidopsis thaliana, suggesting that sucrose was involved in a signalling pathway. In conclusion, we identified potential hormonal candidates for bud outgrowth control by sugar. They are central to future investigations aimed at disentangling the processes that underlie regulation of bud outgrowth by sugar. PMID:25873679

  18. Induction of ectopic taste buds by SHH reveals the competency and plasticity of adult lingual epithelium

    OpenAIRE

    Castillo, David; Seidel, Kerstin; Salcedo, Ernesto; Ahn, Christina; de Sauvage, Frederic J.; Klein, Ophir D.; Linda A Barlow

    2014-01-01

    Taste buds are assemblies of elongated epithelial cells, which are innervated by gustatory nerves that transmit taste information to the brain stem. Taste cells are continuously renewed throughout life via proliferation of epithelial progenitors, but the molecular regulation of this process remains unknown. During embryogenesis, sonic hedgehog (SHH) negatively regulates taste bud patterning, such that inhibition of SHH causes the formation of more and larger taste bud primordia, including in ...

  19. Metabolite changes in conifer buds and needles during forced bud break in Norway spruce (Picea abies and European silver fir (Abies alba

    Directory of Open Access Journals (Sweden)

    Priyanka eDhuli

    2014-12-01

    Full Text Available Environmental changes such as early spring and warm spells induce bud burst and photosynthetic processes in cold-acclimated coniferous trees and consequently, cellular metabolism in overwintering needles and buds. The purpose of the study was to examine metabolism in conifers under forced deacclimation (artificially induced spring by exposing shoots of Picea abies (boreal species and Abies alba (temperate species to a greenhouse environment (22°C, 16/8 h D/N cycle over a nine week period. Each week, we scored bud opening and collected samples for GC/MS–based metabolite profiling. We detected a total of 169 assigned metabolites and 80 identified metabolites, comprising compounds such as mono- and disaccharides, Krebs cycle acids, amino acids, polyols, phenolics and phosphorylated structures. Untargeted multivariate statistical analysis based on PCA and cluster analysis segregated samples by species, tissue type, and stage of tissue deacclimations. Similar patterns of metabolic regulation in both species were observed in buds (amino acids, Krebs cycle acids and needles (hexoses, pentoses, and Krebs cycle acids. Based on correlation of bud opening score with compound levels, distinct metabolites could be associated with bud and shoot development, including amino acids, sugars and acids with known osmolyte function, and secondary metabolites. This study has shed light on how elevated temperature affects metabolism in buds and needles of conifer species during the deacclimation phase, and contributes to the discussion about how phenological characters in conifers may respond to future global warming.

  20. γ-Lactam alkaloids from the flower buds of daylily.

    Science.gov (United States)

    Matsumoto, Takahiro; Nakamura, Seikou; Nakashima, Souichi; Ohta, Tomoe; Yano, Mamiko; Tsujihata, Junichiro; Tsukioka, Junko; Ogawa, Keiko; Fukaya, Masashi; Yoshikawa, Masayuki; Matsuda, Hisashi

    2016-07-01

    Four new alkaloids, hemerocallisamines IV-VII, were isolated from the methanol extract of flower buds of daylily. The chemical structures of the new compounds were elucidated on the basis of chemical and physicochemical evidence. The absolute stereochemistry of the hemerocallisamines IV-VI was elucidated by the application of the modified Mosher's method, HPLC analysis, and optical rotation. In the present study, the isolated alkaloids significantly inhibited the aggregation of Aβ42 in vitro. This is the first report about bioactive alkaloids with a γ-lactam ring from daylily. In addition, isolated nucleosides showed accelerative effects on neurite outgrowth under the non-fasting condition.

  1. Mechanical damage in cotton buds caused by the boll weevil

    Directory of Open Access Journals (Sweden)

    Santos Roseane Cavalcanti

    2003-01-01

    Full Text Available The boll weevil (Anthonomus grandis Boheman causes high levels of bud abscission in cotton plants due to feeding or oviposition punctures. It has been reported that abscission is mainly due to enzymes present in the insect's saliva, but mechanical damage could also contribute to square abscission. The objective of this paper was to undertake an analysis of the morphological damages caused by the insect in cotton squares using microscopy. Anthers and ovules are the main target of boll weevil feeding. The process initiates by perforation of young sepal and petal tissues and proceeds with subsequent alimentation on stamen and ovary leading to abscission of floral structures.

  2. Green Synthesis of Novel Jasmine Bud-Shaped Copper Nanoparticles

    OpenAIRE

    Malathi Sampath; Ramya Vijayan; Ezhilarasu Tamilarasu; Abiraman Tamilselvan; Balasubramanian Sengottuvelan

    2014-01-01

    Novel jasmine bud-shaped copper nanoparticles were synthesized by a green chemical reduction method using polyvinylpyrrolidone (PVP) as a capping agent, L-ascorbic acid (AA) as a reducing agent as well as antioxidant agent, isonicotinic acid hydrazide (INH) as a reducing agent, and water as a solvent at 60–70°C (pH-7) in the presence of air. The UV-Vis absorption maximum obtained is 573 nm. The crystal lattice (fcc) structure of Cu Nps was confirmed by X-ray diffraction (XRD). The novel jasm...

  3. Reconstitution of clathrin-coated pit budding from plasma membranes

    OpenAIRE

    1991-01-01

    Receptor-mediated endocytosis begins with the binding of ligand to receptors in clathrin-coated pits followed by the budding of the pits away from the membrane. We have successfully reconstituted this sequence in vitro. Highly purified plasma membranes labeled with gold were obtained by incubating cells in the presence of anti-LDL receptor IgG-gold at 4 degrees C, attaching the labeled cells to a poly-L-lysine- coated substratum at 4 degrees C and then gently sonicating them to remove everyth...

  4. Qualitative and quantitative differences between taste buds of the rat and mouse

    Directory of Open Access Journals (Sweden)

    Ma Huazhi

    2007-01-01

    Full Text Available Abstract Background Numerous electrophysiological, ultrastructural, and immunocytochemical studies on rodent taste buds have been carried out on rat taste buds. In recent years, however, the mouse has become the species of choice for molecular and other studies on sensory transduction in taste buds. Do rat and mouse taste buds have the same cell types, sensory transduction markers and synaptic proteins? In the present study we have used antisera directed against PLCβ2, α-gustducin, serotonin (5-HT, PGP 9.5 and synaptobrevin-2 to determine the percentages of taste cells expressing these markers in taste buds in both rodent species. We also determined the numbers of taste cells in the taste buds as well as taste bud volume. Results There are significant differences (p 3 is smaller than a rat taste bud (64,200 μm3. The numerical density of taste cells in mouse circumvallate taste buds (2.1 cells/1000 μm3 is significantly higher than that in the rat (1.2 cells/1000 μm3. Conclusion These results suggest that rats and mice differ significantly in the percentages of taste cells expressing signaling molecules. We speculate that these observed dissimilarities may reflect differences in their gustatory processing.

  5. Bud protection: a key trait for species sorting in a forest-savanna mosaic.

    Science.gov (United States)

    Charles-Dominique, Tristan; Beckett, Heath; Midgley, Guy F; Bond, William J

    2015-09-01

    Contrasting fire regimes maintain patch mosaics of savanna, thicket and forest biomes in many African subtropical landscapes. Species dominating each biome are thus expected to display distinct fire-related traits, commonly thought to be bark related. Recent Australian savanna research suggests that bud position, not bark protection alone, determines fire resilience via resprouting. We tested first how bud position influences resprouting ability in 17 tree species. We then compared the effect of both bark-related protection and bud position on the distribution of 63 tree species in 253 transects in all three biomes. Tree species with buds positioned deep under bark had a higher proportion of post-fire aboveground shoot resprouting. Species with low bud protection occurred in fire-prone biomes only if they could root-sucker. The effect of bud protection was supported by a good relationship between species bud protection and distribution across a gradient of fire frequency. Bud protection and high bark production are required to survive frequent fires in savanna. Forests are fire refugia hosting species with little or no bud protection and thin bark. Root-suckering species occur in the three biomes, suggesting that fire is not the only factor filtering this functional type. PMID:25856385

  6. Analysis of Essential Oils from Flower-buds, Leaves and Stems of Filipendula palmata (Pall.) Maxim.

    Institute of Scientific and Technical Information of China (English)

    WU Yin; MENG Xiang-ying; LIU Xiao-hua; BAO Yong-li; WANG Shu-ping; LI Yu-xin

    2005-01-01

    The essential oils of the flower-buds, leaves and stems of Filipendula palmata (Pall.) Maxim. were analyzed by GC and GC-MS. Thirty-three, forty-seven and forty-seven compounds in the flower-buds, leaves and stems were identified, respectively. Methyl salicylate exists in a great amount which is up to 70. 10% in the flower-buds. Its amount is also high in other two parts. The data obtained show that it may be one of the main natural mosquito-expelling and pain-alleviating components in the three parts. The flower-buds are the main active part with the mosquito-expelling function.

  7. Quick detection of Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae) in chestnut dormant buds by nested PCR.

    Science.gov (United States)

    Sartor, C; Marinoni, D Torello; Quacchia, A; Botta, R

    2012-06-01

    Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae) develops in chestnut buds that remain asymptomatic from oviposition (June-July) until budburst; it is, thus, easily spread by plant material used in propagation. Therefore, it is particularly interesting to identify infested plant batches before their movement. Unfortunately, a non-destructive method for checking buds has not yet been developed, and the only technique available is the screening of a bud sample. The visual investigation is long and requires highly skilled and trained staff. The purpose of this work was to set up an effective and fast method able to identify the presence of first instar larvae of D. kuriphilus in a large number of chestnut buds by PCR. Four primer pairs were designed on nuclear and mitochondrial sequences of a set of seven gall wasp taxa and tested on five different cynipid's DNA. Nested diagnostic PCR was carried out on DNA extracted from samples of 2 g buds simulating four levels of infestation (larvae were added to uninfested buds); 320 bp amplicon of 28S sequence was chosen as a marker to detect one larva out of 2 g buds. The method showed a potential efficiency of 5000 to 15,000 buds per week, depending on bud size.

  8. A computational clonal analysis of the developing mouse limb bud.

    Directory of Open Access Journals (Sweden)

    Luciano Marcon

    Full Text Available A comprehensive spatio-temporal description of the tissue movements underlying organogenesis would be an extremely useful resource to developmental biology. Clonal analysis and fate mappings are popular experiments to study tissue movement during morphogenesis. Such experiments allow cell populations to be labeled at an early stage of development and to follow their spatial evolution over time. However, disentangling the cumulative effects of the multiple events responsible for the expansion of the labeled cell population is not always straightforward. To overcome this problem, we develop a novel computational method that combines accurate quantification of 2D limb bud morphologies and growth modeling to analyze mouse clonal data of early limb development. Firstly, we explore various tissue movements that match experimental limb bud shape changes. Secondly, by comparing computational clones with newly generated mouse clonal data we are able to choose and characterize the tissue movement map that better matches experimental data. Our computational analysis produces for the first time a two dimensional model of limb growth based on experimental data that can be used to better characterize limb tissue movement in space and time. The model shows that the distribution and shapes of clones can be described as a combination of anisotropic growth with isotropic cell mixing, without the need for lineage compartmentalization along the AP and PD axis. Lastly, we show that this comprehensive description can be used to reassess spatio-temporal gene regulations taking tissue movement into account and to investigate PD patterning hypothesis.

  9. Green Synthesis of Novel Jasmine Bud-Shaped Copper Nanoparticles

    Directory of Open Access Journals (Sweden)

    Malathi Sampath

    2014-01-01

    Full Text Available Novel jasmine bud-shaped copper nanoparticles were synthesized by a green chemical reduction method using polyvinylpyrrolidone (PVP as a capping agent, L-ascorbic acid (AA as a reducing agent as well as antioxidant agent, isonicotinic acid hydrazide (INH as a reducing agent, and water as a solvent at 60–70°C (pH-7 in the presence of air. The UV-Vis absorption maximum obtained is 573 nm. The crystal lattice (fcc structure of Cu Nps was confirmed by X-ray diffraction (XRD. The novel jasmine bud shape was visualized in a transmission electron microscope (TEM. The height of single copper nanobud was 6.41 nm as measured by atomic force microscope (AFM. The average particle size 6.95 nm is obtained by XRD results. Antibacterial activity of the Cu nanobuds was evaluated by testing against Gram-negative (Escherichia coli and Gram-positive (Staphylococcus aureus bacteria.

  10. BudBurst Buddies: Introducing Young Citizen Scientists to Plants and Environmental Change

    Science.gov (United States)

    Ward, D.; Gardiner, L. S.; Henderson, S.

    2011-12-01

    As part of Project BudBurst, the BudBurst Buddies recently moved to the National Ecological Network (NEON) as part of its Education and Public Engagement efforts. The BudBurst Buddies (www.budburstbuddies.org) were created to engage elementary school age children in the science of observing plants and the timing of phenological (life cycle) events. BudBurst Buddies is a part of the Project BudBurst national citizen science initiative (www.budburst.org), which allows individuals to engage in the scientific process, contributing to a better understanding of climate change while increasing public awareness of phenology and the impacts of climate change on plants. As a first step towards engaging the next generation of citizen scientists, BudBurst Buddies provides the opportunity for children to gain experience with scientific research and increases awareness of how plants change throughout the year. Hundreds of young students have participated in the inaugural year of BudBurst Buddies. Children can participate in BudBurst Buddies on their own, with their families, or in formal or informal education settings. The program was recently highlighted by education staff at the New York Hall of Science and numerous classrooms have been implementing this resource as part of their curriculum. Each child who participates creates a journal about a plant of his or her choosing, makes observations of the plant over the growing season and submits findings online, earning an official BudBurst Buddies certificate. An online storybook for kids tells how two children, Lily and Sage, observed plants in their neighborhood and became BudBurst Buddies. This presentation will provide an overview of the BudBurst Buddies resources including a new implementation guide and will also share feedback from the first year of implementation.

  11. Vector sequences - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available Budding yeast cDNA sequencing project Vector sequences Data detail Data name Vector sequences Description of data contents Vector seq...wnload License Update History of This Database Site Policy | Contact Us Vector sequences - Budding yeast cDNA sequencing project | LSDB Archive ... ...uences used for sequencing. Multi FASTA format. 7 entries. Data file File name: vec

  12. Uptake and distribution of 32P in the budded and self-rooted grape varieties

    International Nuclear Information System (INIS)

    In the self-rooted and budded varieties of grape (Vitis vinifera L.), the total P and 32P contents were high in 'Anabee-Shahi', but low in in 'Muscat'. The growing shoots contained more P than old stems and roots in all the varieties. In the budded plants, 'Kali Sahebi' scion budded on 'Anab-e-Shani showed the maximum 32P and total P in the shoots, but 'Muscat' scion budded on 'Anab-e-Shahi' accumulated more P in the roots and very low 32P in the growing shoots. Auto-radiographs of shoots also showed that 'Kali Sahebi' budded on 'Anab-e-Shani' rootstock accumulated more 32P in the shoots. (author)

  13. Cell kinetic study on the relation between irradiation hypogeusia and taste buds in rats

    Energy Technology Data Exchange (ETDEWEB)

    Kubota, Hideharu; Furumoto, Keiichi [Nippon Dental Univ., Tokyo (Japan)

    1998-12-01

    The present study was designed to elucidate the mechanism of hypogeusia caused by irradiation. X-ray treatment at 10 Gy or 20 Gy was given to the maxillofacial region including the tongue in rats, and the involvement of taste bud for hypogeusia was investigated. In addition, cytological kinetics were immunohistologically studied using bromodeoxyuridine in the taste bud and in the lingual mucosal epithelium. The following results were obtained: In the 10 Gy group, the number of taste bud become less after the exposure, but no hypogeusia was observed during the experimental period. In the 20 Gy group, any labeled taste bud was not observed on the 7th day, and all taste buds disappeared by the 10th day. In the lingual mucosal epithelium, the number of basal cells decreased to the minimum, and the body weight and total water intake decreased coincidently in the 20 Gy group, which were few in the 10 Gy group. (author)

  14. BudBurst Buddies: A New Tool for Engaging the Youngest Citizen Scientists

    Science.gov (United States)

    Gardiner, L. S.; Henderson, S.; Ward, D.

    2010-12-01

    BudBurst Buddies (www.budburstbuddies.org) introduces elementary school age children to the science of observing plants and the timing of phenological (life cycle) events. BudBurst Buddies is a new part of the Project BudBurst national citizen science initiative (www.budburst.org), which allows individuals to engage in the scientific process, contributing to a better understanding of climate change while increasing public awareness of phenology and the impacts of climate change on plants. As a first step towards engaging the next generation of citizen scientists, BudBurst Buddies provides the opportunity for children to gain experience with scientific research and increases awareness of how plants change throughout the year. Children can participate in BudBurst Buddies on their own, with their families, or in formal or informal education settings. Each child who participates creates a journal about a plant of his or her choosing, makes observations of the plant over the growing season and submits findings online, earning an official BudBurst Buddies certificate. An online storybook for kids tells how two children, Lily and Sage, observed plants in their neighborhood and became BudBurst Buddies. This presentation will provide an overview of the BudBurst Buddies newly developed resources. BudBurst Buddies is a part of Project BudBurst, a national citizen science program coordinated by the National Ecological Observatory Network (NEON) and the Chicago Botanic Garden. Funding for this resource was provided by NEON, NSF, NASA, and the National Geographic Education Foundation.

  15. Breaking-bud pollination: a new pollination process in partially opened flowers by small bees.

    Science.gov (United States)

    Yamaji, Futa; Ohsawa, Takeshi A

    2015-09-01

    Plant-pollinator interactions have usually been researched in flowers that have fully opened. However, some pollinators can visit flowers before full opening and contribute to fruit and seed sets. In this paper, we researched the pollination biology of flowers just starting to open in four field experiments. We observed the insect visitors to Lycoris sanguinea var. sanguinea for 3 years at five sites. These observations revealed that only small bees, Lasioglossum japonicum, often entered through tiny spaces between the tepals of 'breaking buds' (i.e. partially opened flowers) and collected pollen. We hypothesized that they can pollinate this species at the breaking-bud stage, when the stigma is located near the anthers. To measure the pollination effect of small bees at the breaking-bud stage, we bagged several breaking buds after small bees had visited them and examined whether these buds were pollinated. In bagging experiments, 30% of the breaking buds set fruit and seeds. Fruit-set ratios of the breaking buds did not differ significantly from those of the fully opened flowers, which had been visited by several insect species. We also counted the pollen grain numbers on the body of L. japonicum and on the anthers of randomly-selected and manipulated flowers. These experiments revealed that all of the captured bees had some pollen of target plants and that L. japonicum collected most of the pollen grains at the breaking-bud stage. Our results showed that the new pollination process, breaking-bud pollination, happened in breaking buds by L. japonicum, although there is no evidence to reveal that this is the most effective pollination method for L. sanguinea var. sanguinea. In principle, this new pollination process can occur in other flowering plants and our results are a major contribution to studies of plant-pollinator interactions.

  16. Novel Features of the Prenatal Horn Bud Development in Cattle (Bos taurus.

    Directory of Open Access Journals (Sweden)

    Dominique Judith Wiener

    Full Text Available Whereas the genetic background of horn growth in cattle has been studied extensively, little is known about the morphological changes in the developing fetal horn bud. In this study we histologically analyzed the development of horn buds of bovine fetuses between ~70 and ~268 days of pregnancy and compared them with biopsies taken from the frontal skin of the same fetuses. In addition we compared the samples from the wild type (horned fetuses with samples taken from the horn bud region of age-matched genetically hornless (polled fetuses. In summary, the horn bud with multiple layers of vacuolated keratinocytes is histologically visible early in fetal life already at around day 70 of gestation and can be easily differentiated from the much thinner epidermis of the frontal skin. However, at the gestation day (gd 212 the epidermis above the horn bud shows a similar morphology to the epidermis of the frontal skin and the outstanding layers of vacuolated keratinocytes have disappeared. Immature hair follicles are seen in the frontal skin at gd 115 whereas hair follicles below the horn bud are not present until gd 155. Interestingly, thick nerve bundles appear in the dermis below the horn bud at gd 115. These nerve fibers grow in size over time and are prominent shortly before birth. Prominent nerve bundles are not present in the frontal skin of wild type or in polled fetuses at any time, indicating that the horn bud is a very sensitive area. The samples from the horn bud region from polled fetuses are histologically equivalent to samples taken from the frontal skin in horned species. This is the first study that presents unique histological data on bovine prenatal horn bud differentiation at different developmental stages which creates knowledge for a better understanding of recent molecular findings.

  17. Guillaume Budé, l’humaniste et le prince

    Directory of Open Access Journals (Sweden)

    Sylvie Le Clech-Charton

    2009-09-01

    Full Text Available Grande figure de la Renaissance des lettres et des arts en France, tout à la fois écrivain, traducteur, ambassadeur, créateur du dépôt légal et fondateur du Collège de France, maître de la librairie du roi à Fontainebleau, Guillaume Budé (1468-1540 est essentiellement connu pour le rôle de conseiller politique et culturel qu’il joua auprès de François Ier, dont il fut le secrétaire. Il a été surtout étudié du point de vue de sa production littéraire savante, mais non sous l’angle de son mili...

  18. Dynamical Analysis of Protein Regulatory Network in Budding Yeast Nucleus

    Institute of Scientific and Technical Information of China (English)

    LI Fang-Ting; JIA Xun

    2006-01-01

    @@ Recent progresses in the protein regulatory network of budding yeast Saccharomyces cerevisiae have provided a global picture of its protein network for further dynamical research. We simplify and modularize the protein regulatory networks in yeast nucleus, and study the dynamical properties of the core 37-node network by a Boolean network model, especially the evolution steps and final fixed points. Our simulation results show that the number of fixed points N(k) for a given size of the attraction basin k obeys a power-law distribution N(k)∝k-2.024. The yeast network is more similar to a scale-free network than a random network in the above dynamical properties.

  19. The physics of lipid droplet nucleation, growth and budding.

    Science.gov (United States)

    Thiam, Abdou Rachid; Forêt, Lionel

    2016-08-01

    Lipid droplets (LDs) are intracellular oil-in-water emulsion droplets, covered by a phospholipid monolayer and mainly present in the cytosol. Despite their important role in cellular metabolism and growing number of newly identified functions, LD formation mechanism from the endoplasmic reticulum remains poorly understood. To form a LD, the oil molecules synthesized in the ER accumulate between the monolayer leaflets and induce deformation of the membrane. This formation process works through three steps: nucleation, growth and budding, exactly as in phase separation and dewetting phenomena. These steps involve sequential biophysical membrane remodeling mechanisms for which we present basic tools of statistical physics, membrane biophysics, and soft matter science underlying them. We aim to highlight relevant factors that could control LD formation size, site and number through this physics description. An emphasis will be given to a currently underestimated contribution of the molecular interactions between lipids to favor an energetically costless mechanism of LD formation. PMID:27131867

  20. Carbohydrate concentrations and freezing stress resistance of silver birch buds grown under elevated temperature and ozone.

    Science.gov (United States)

    Riikonen, Johanna; Kontunen-Soppela, Sari; Vapaavuori, Elina; Tervahauta, Arja; Tuomainen, Marjo; Oksanen, Elina

    2013-03-01

    The effects of slightly elevated temperature (+0.8 °C), ozone (O3) concentration (1.3 × ambient O3 concentration) and their combination on over-wintering buds of Betula pendula Roth were studied after two growing seasons of exposure in the field. Carbohydrate concentrations, freezing stress resistance (FSR), bud dry weight to fresh weight ratio, and transcript levels of cytochrome oxidase (COX), alternative oxidase (AOX) and dehydrin (LTI36) genes were studied in two clones (clones 12 and 25) in December. Elevated temperature increased the bud dry weight to fresh weight ratio and the ratio of raffinose family oligosaccharides to sucrose and the transcript levels of the dehydrin (LTI36) gene (in clone 12 only), but did not alter the FSR of the buds. Genotype-specific alterations in carbohydrate metabolism were found in the buds grown under elevated O3. The treatments did not significantly affect the transcript level of the COX or AOX genes. No clear pattern of an interactive effect between elevated temperature and O3 concentration was found. According to these data, the increase in autumnal temperatures and slightly increasing O3 concentrations do not increase the risk for freeze-induced damage in winter in silver birch buds, although some alterations in bud physiology occur.

  1. Automated quantification of budding Saccharomyces cerevisiae using a novel image cytometry method.

    Science.gov (United States)

    Laverty, Daniel J; Kury, Alexandria L; Kuksin, Dmitry; Pirani, Alnoor; Flanagan, Kevin; Chan, Leo Li-Ying

    2013-06-01

    The measurements of concentration, viability, and budding percentages of Saccharomyces cerevisiae are performed on a routine basis in the brewing and biofuel industries. Generation of these parameters is of great importance in a manufacturing setting, where they can aid in the estimation of product quality, quantity, and fermentation time of the manufacturing process. Specifically, budding percentages can be used to estimate the reproduction rate of yeast populations, which directly correlates with metabolism of polysaccharides and bioethanol production, and can be monitored to maximize production of bioethanol during fermentation. The traditional method involves manual counting using a hemacytometer, but this is time-consuming and prone to human error. In this study, we developed a novel automated method for the quantification of yeast budding percentages using Cellometer image cytometry. The automated method utilizes a dual-fluorescent nucleic acid dye to specifically stain live cells for imaging analysis of unique morphological characteristics of budding yeast. In addition, cell cycle analysis is performed as an alternative method for budding analysis. We were able to show comparable yeast budding percentages between manual and automated counting, as well as cell cycle analysis. The automated image cytometry method is used to analyze and characterize corn mash samples directly from fermenters during standard fermentation. Since concentration, viability, and budding percentages can be obtained simultaneously, the automated method can be integrated into the fermentation quality assurance protocol, which may improve the quality and efficiency of beer and bioethanol production processes.

  2. Auxin dynamics after decapitation are not correlated with the initial growth of axillary buds.

    Science.gov (United States)

    Morris, Suzanne E; Cox, Marjolein C H; Ross, John J; Krisantini, Santi; Beveridge, Christine A

    2005-07-01

    One of the first and most enduring roles identified for the plant hormone auxin is the mediation of apical dominance. Many reports have claimed that reduced stem indole-3-acetic acid (IAA) levels and/or reduced basipetal IAA transport directly or indirectly initiate bud growth in decapitated plants. We have tested whether auxin inhibits the initial stage of bud release, or subsequent stages, in garden pea (Pisum sativum) by providing a rigorous examination of the dynamics of auxin level, auxin transport, and axillary bud growth. We demonstrate that after decapitation, initial bud growth occurs prior to changes in IAA level or transport in surrounding stem tissue and is not prevented by an acropetal supply of exogenous auxin. We also show that auxin transport inhibitors cause a similar auxin depletion as decapitation, but do not stimulate bud growth within our experimental time-frame. These results indicate that decapitation may trigger initial bud growth via an auxin-independent mechanism. We propose that auxin operates after this initial stage, mediating apical dominance via autoregulation of buds that are already in transition toward sustained growth. PMID:15965021

  3. Phenotypic plasticity, QTL mapping and genomic characterization of bud set in black poplar

    Directory of Open Access Journals (Sweden)

    Fabbrini Francesco

    2012-04-01

    Full Text Available Abstract Background The genetic control of important adaptive traits, such as bud set, is still poorly understood in most forest trees species. Poplar is an ideal model tree to study bud set because of its indeterminate shoot growth. Thus, a full-sib family derived from an intraspecific cross of P. nigra with 162 clonally replicated progeny was used to assess the phenotypic plasticity and genetic variation of bud set in two sites of contrasting environmental conditions. Results Six crucial phenological stages of bud set were scored. Night length appeared to be the most important signal triggering the onset of growth cessation. Nevertheless, the effect of other environmental factors, such as temperature, increased during the process. Moreover, a considerable role of genotype × environment (G × E interaction was found in all phenological stages with the lowest temperature appearing to influence the sensitivity of the most plastic genotypes. Descriptors of growth cessation and bud onset explained the largest part of phenotypic variation of the entire process. Quantitative trait loci (QTL for these traits were detected. For the four selected traits (the onset of growth cessation (date2.5, the transition from shoot to bud (date1.5, the duration of bud formation (subproc1 and bud maturation (subproc2 eight and sixteen QTL were mapped on the maternal and paternal map, respectively. The identified QTL, each one characterized by small or modest effect, highlighted the complex nature of traits involved in bud set process. Comparison between map location of QTL and P. trichocarpa genome sequence allowed the identification of 13 gene models, 67 bud set-related expressional and six functional candidate genes (CGs. These CGs are functionally related to relevant biological processes, environmental sensing, signaling, and cell growth and development. Some strong QTL had no obvious CGs, and hold great promise to identify unknown genes that affect bud set

  4. Voltage-gated sodium channels in taste bud cells

    Directory of Open Access Journals (Sweden)

    Williams Mark E

    2009-03-01

    Full Text Available Abstract Background Taste bud cells transmit information regarding the contents of food from taste receptors embedded in apical microvilli to gustatory nerve fibers innervating basolateral membranes. In particular, taste cells depolarize, activate voltage-gated sodium channels, and fire action potentials in response to tastants. Initial cell depolarization is attributable to sodium influx through TRPM5 in sweet, bitter, and umami cells and an undetermined cation influx through an ion channel in sour cells expressing PKD2L1, a candidate sour taste receptor. The molecular identity of the voltage-gated sodium channels that sense depolarizing signals and subsequently initiate action potentials coding taste information to gustatory nerve fibers is unknown. Results We describe the molecular and histological expression profiles of cation channels involved in electrical signal transmission from apical to basolateral membrane domains. TRPM5 was positioned immediately beneath tight junctions to receive calcium signals originating from sweet, bitter, and umami receptor activation, while PKD2L1 was positioned at the taste pore. Using mouse taste bud and lingual epithelial cells collected by laser capture microdissection, SCN2A, SCN3A, and SCN9A voltage-gated sodium channel transcripts were expressed in taste tissue. SCN2A, SCN3A, and SCN9A were expressed beneath tight junctions in subsets of taste cells. SCN3A and SCN9A were expressed in TRPM5 cells, while SCN2A was expressed in TRPM5 and PKD2L1 cells. HCN4, a gene previously implicated in sour taste, was expressed in PKD2L1 cells and localized to cell processes beneath the taste pore. Conclusion SCN2A, SCN3A and SCN9A voltage-gated sodium channels are positioned to sense initial depolarizing signals stemming from taste receptor activation and initiate taste cell action potentials. SCN2A, SCN3A and SCN9A gene products likely account for the tetrodotoxin-sensitive sodium currents in taste receptor cells.

  5. Tanshinones extend chronological lifespan in budding yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Wu, Ziyun; Song, Lixia; Liu, Shao Quan; Huang, Dejian

    2014-10-01

    Natural products with anti-aging property have drawn great attention recently but examples of such compounds are exceedingly scarce. By applying a high-throughput assay based on yeast chronological lifespan measurement, we screened the anti-aging activity of 144 botanical materials and found that dried roots of Salvia miltiorrhiza Bunge have significant anti-aging activity. Tanshinones isolated from the plant including cryptotanshione, tanshinone I, and tanshinone IIa, are the active components. Among them, cryptotanshinone can greatly extend the budding yeast Saccharomyces cerevisiae chronological lifespan (up to 2.5 times) in a dose- and the-time-of-addition-dependent manner at nanomolar concentrations without disruption of cell growth. We demonstrate that cryptotanshinone prolong chronological lifespan via a nutrient-dependent regime, especially essential amino acid sensing, and three conserved protein kinases Tor1, Sch9, and Gcn2 are required for cryptotanshinone-induced lifespan extension. In addition, cryptotanshinone significantly increases the lifespan of SOD2-deleted mutants. Altogether, those data suggest that cryptotanshinone might be involved in the regulation of, Tor1, Sch9, Gcn2, and Sod2, these highly conserved longevity proteins modulated by nutrients from yeast to humans.

  6. Regulation of the divalent metal ion transporter via membrane budding

    Science.gov (United States)

    Mackenzie, KimberlyD; Foot, Natalie J; Anand, Sushma; Dalton, Hazel E; Chaudhary, Natasha; Collins, Brett M; Mathivanan, Suresh; Kumar, Sharad

    2016-01-01

    The release of extracellular vesicles (EVs) is important for both normal physiology and disease. However, a basic understanding of the targeting of EV cargoes, composition and mechanism of release is lacking. Here we present evidence that the divalent metal ion transporter (DMT1) is unexpectedly regulated through release in EVs. This process involves the Nedd4-2 ubiquitin ligase, and the adaptor proteins Arrdc1 and Arrdc4 via different budding mechanisms. We show that mouse gut explants release endogenous DMT1 in EVs. Although we observed no change in the relative amount of DMT1 released in EVs from gut explants in Arrdc1 or Arrdc4 deficient mice, the extent of EVs released was significantly reduced indicating an adaptor role in biogenesis. Furthermore, using Arrdc1 or Arrdc4 knockout mouse embryonic fibroblasts, we show that both Arrdc1 and Arrdc4 are non-redundant positive regulators of EV release. Our results suggest that DMT1 release from the plasma membrane into EVs may represent a novel mechanism for the maintenance of iron homeostasis, which may also be important for the regulation of other membrane proteins. PMID:27462458

  7. Asymmetric nucleosomes flank promoters in the budding yeast genome.

    Science.gov (United States)

    Ramachandran, Srinivas; Zentner, Gabriel E; Henikoff, Steven

    2015-03-01

    Nucleosomes in active chromatin are dynamic, but whether they have distinct structural conformations is unknown. To identify nucleosomes with alternative structures genome-wide, we used H4S47C-anchored cleavage mapping, which revealed that 5% of budding yeast (Saccharomyces cerevisiae) nucleosome positions have asymmetric histone-DNA interactions. These asymmetric interactions are enriched at nucleosome positions that flank promoters. Micrococcal nuclease (MNase) sequence-based profiles of asymmetric nucleosome positions revealed a corresponding asymmetry in MNase protection near the dyad axis, suggesting that the loss of DNA contacts around H4S47 is accompanied by protection of the DNA from MNase. Chromatin immunoprecipitation mapping of selected nucleosome remodelers indicated that asymmetric nucleosomes are bound by the RSC chromatin remodeling complex, which is required for maintaining nucleosomes at asymmetric positions. These results imply that the asymmetric nucleosome-RSC complex is a metastable intermediate representing partial unwrapping and protection of nucleosomal DNA on one side of the dyad axis during chromatin remodeling.

  8. Timing robustness in the budding and fission yeast cell cycles.

    KAUST Repository

    Mangla, Karan

    2010-02-01

    Robustness of biological models has emerged as an important principle in systems biology. Many past analyses of Boolean models update all pending changes in signals simultaneously (i.e., synchronously), making it impossible to consider robustness to variations in timing that result from noise and different environmental conditions. We checked previously published mathematical models of the cell cycles of budding and fission yeast for robustness to timing variations by constructing Boolean models and analyzing them using model-checking software for the property of speed independence. Surprisingly, the models are nearly, but not totally, speed-independent. In some cases, examination of timing problems discovered in the analysis exposes apparent inaccuracies in the model. Biologically justified revisions to the model eliminate the timing problems. Furthermore, in silico random mutations in the regulatory interactions of a speed-independent Boolean model are shown to be unlikely to preserve speed independence, even in models that are otherwise functional, providing evidence for selection pressure to maintain timing robustness. Multiple cell cycle models exhibit strong robustness to timing variation, apparently due to evolutionary pressure. Thus, timing robustness can be a basis for generating testable hypotheses and can focus attention on aspects of a model that may need refinement.

  9. Molecular cytotoxicity mechanisms of allyl alcohol (acrolein) in budding yeast.

    Science.gov (United States)

    Golla, Upendarrao; Bandi, Goutham; Tomar, Raghuvir S

    2015-06-15

    Allyl alcohol (AA) is one of the environmental pollutants used as a herbicide and industrial chemical. AA undergoes enzymatic oxidation in vivo to form Acrolein (Acr), a highly reactive and ubiquitous environmental toxicant. The exposure to AA/Acr has detrimental effects on cells and is highly fatal. In corroboration to the current literature describing AA/Acr toxicity, this study aimed to investigate the molecular cytotoxicity mechanisms of AA/Acr using budding yeast as a eukaryotic model organism. Genome-wide transcriptome analysis of cells treated with a sublethal dose of AA (0.4 mM) showed differential regulation of approximately 30% of the yeast genome. Functional enrichment analysis of the AA transcriptome revealed that genes belong to diverse cellular processes including the cell cycle, DNA damage repair, metal homeostasis, stress response genes, ribosomal biogenesis, metabolism, meiosis, ubiquitination, cell morphogenesis, and transport. Moreover, we have identified novel molecular targets of AA/Acr through genetic screening, which belongs to oxidative stress, DNA damage repair, iron homeostasis, and cell wall integrity. This study also demonstrated the epigenetic basis of AA/Acr toxicity mediated through histone tails and chromatin modifiers. Interestingly, our study disclosed the use of pyrazole and ethanol as probable antidotes for AA intoxication. For the first time, this study also demonstrated the reproductive toxicity of AA/Acr using the yeast gametogenesis (spermatogenesis) model. Altogether, this study unravels the molecular mechanisms of AA/Acr cytotoxicity and facilitates the prediction of biomarkers for toxicity assessment and therapeutic approaches. PMID:25919230

  10. Automatic nuclear bud detection using ellipse fitting, moving sticks or top-hat transformation.

    Science.gov (United States)

    Zhang, C; Sun, C; Vallotton, P; Fenech, M; Pham, T D

    2013-11-01

    Micronucleus assays are extensively used by biologists to assess genotoxicity and to monitor human exposure to genotoxic materials. As recent studies suggested that nuclear buds can be a new source of micronuclei formed in interphase, the quantification of nuclear buds, which are micronucleus like objects that are attached to the nuclei in interphase, in normal and control group is needed. Three automatic nuclear bud detection algorithms fit for different situations are proposed in this paper. One is based on ellipse fitting, one is based on a stick model and the other is based on the top-hat transform. Comparison of the three methods is also given in this paper. Experimental results showed that the proposed algorithms are all effective and efficient for nuclear bud detection. PMID:23961938

  11. Field and laboratory investigations of budding in the tetillid sponge Cinachyrella cavernosa

    Digital Repository Service at National Institute of Oceanography (India)

    Singh, A.; Thakur, N.L.

    C. australiensis, T. citrina and T. aurantium. The compact structure of these buds can be speculated to be an adaptive strategy to reduce the cost of the budding process, thus making it energetically favored in a stressful environment... Author for correspondence. Email address: thakurn@nio.org 2 Marine invertebrates such as corals, sponges, sea urchins, sea cucumbers, sea anemones and barnacles can adopt different reproductive strategies (e.g., asexual and /or sexual...

  12. Electron microscopic observation of hepatitis B virus budding from hepatocytes into bile canaliculi.

    OpenAIRE

    Ymadada, Gotaro; Sakamoto,Yuji; Mizuno, Motowo; Kobayashi, Toshinari; Nagashima,Hideo

    1980-01-01

    In electron microscopic observation of a liver biopsy obtained from a hepatitis B surface antigen-positive patient, noncoated core particles were occasionally seen budding into the hepatocytic cisterni and many Dane particles were found in the pericanalicular vesicles of hepatocytes. Noncoated core particles were also localized in clusters within the bleb of microvilli. There were some core particles being protruded from microvilli into the lumen of bile canaliculi by budding. These findings ...

  13. Sea buckthorn bud extract displays activity against cell-cultured Influenza virus

    OpenAIRE

    TORELLI, A.; GIANCHECCHI, E.; PICCIRELLA, S.; MANENTI, A.; Piccini, G.; LLORENTE PASTOR, E.; CANOVI, B.; MONTOMOLI, E.

    2015-01-01

    Summary Introduction. Vaccines and antiviral drugs are the most widely used methods of preventing or treating Influenza virus infection. The role of sea buckthorn (SBT) bud dry extract as a natural antiviral drug against Influenza was investigated. Methods. Influenza virus was cultured in the MDCK cell line, with or without SBT bud extract, and virus growth was assessed by HA and TCID50 virus titration in terms of cytopathic effect on cells. Several concentrations of extract were tested, the ...

  14. Marketingová komunikace destinace České Budějovice

    OpenAIRE

    MASARYKOVÁ, Lenka

    2013-01-01

    The thesis deals with an analysis of marketing communication instruments in the destination of České Budějovice. Pursuant of an analysis the current instruments of marketing communication are identified and the efficiency is evaluated. The one part of the thesis is also the suggestion of on optimal communication mix and the measures with contribute to the next development of marketing communication of the destination České Budějovice.

  15. Gustatory stimuli representing different perceptual qualities elicit distinct patterns of neuropeptide secretion from taste buds

    OpenAIRE

    Geraedts, Maartje C. P.; Munger, Steven D.

    2013-01-01

    Taste stimuli that evoke different perceptual qualities (e.g., sweet, umami, bitter, sour, salty) are detected by dedicated subpopulations of taste bud cells that employ distinct combinations of sensory receptors and transduction molecules. Here, we report that taste stimuli also elicit unique patterns of neuropeptide secretion from taste buds that are correlated with those perceptual qualities. We measured tastant-dependent secretion of glucagon-like peptide-1 (GLP-1), glucagon and neuropept...

  16. Association of Dermatological Conditions of External Ear with the Use of Cotton Buds

    Directory of Open Access Journals (Sweden)

    Salahuddin Ahmed

    2014-09-01

    Full Text Available Background: The habit of cleaning the external auditory canal with cotton buds is a common practice of the masses. It has strong association with neurodermatitis and contact dermatitis of the external ear. It is also associated with acute otitis externa, rupture of tympanic membrane causing bleeding and temporary hearing loss in some cases. In many cases the injury will heal but damage to minuscule bones deep inside the ear can cause permanent deafness. Objective: The objective of this study was to determine the association of dermatological condition of external ear with the use of cotton buds. Materials and Methods: This case control study was done from January to October 2012 in the Ear Nose Throat Department of Pakistan Level III Hospital, Darfur, Sudan. Sixty seven patients with dermatological diseases of external ear were cases and 83 subjects without dermatological diseases of external ear were selected as controls. Results: Among 67 cases, 58 were cotton bud users and among 83 controls only 29 were cotton bud users. Different types of dermatological diseases were neurodermatitis (34.32%, otitis externa (28.36%, contact dermatitis (26.87% and wax impaction (8.95%. Ninety three percent of cotton bud users were ignorant of harmful effects of this bad habit. Conclusion: There is a strong association of dermatological diseases of external ear with the use of cotton bud which should be discouraged by fortifying the warning by manufacturers and health education at various educational levels.

  17. Structural Characterization of Ginsenosides from Flower Buds of Panax ginseng by RRLC-Q-TOF MS.

    Science.gov (United States)

    Wu, Wei; Lu, Ziyan; Teng, Yaran; Guo, Yingying; Liu, Shuying

    2016-02-01

    Ginseng flower bud as a part of Panax ginseng has received much attention as a valuable functional food with medicinal potential. A few studies focused on systematic and comprehensive studies on its major ingredients. This study aims to rapidly characterize ginsenosides in ginseng flower buds and provide scientific basis for developing functional food, exploiting pharmaceutical effects and making full use of ginseng resources. A rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (RRLC-Q-TOF-MS) method was developed for rapid qualitative and quantitative analysis of ginsenosides in ginseng flower buds. The compounds were identified by comparing retention time of the reference standards, accurate mass measurement and the fragment ions obtained from RRLC-Q-TOF-MS/MS analyses. A total of 14 kinds of ginsenosides were identified and 5 kinds of malonyl-ginsenosides were first tentatively identified in ginseng flower buds. Ten kinds of main ginsenosides were quantitatively analyzed. The developed RRLC-Q-TOF-MS method was demonstrated as an effective analytical means for rapid characterization of the ginsenosides in flower buds of P. ginseng. The research result is valuable for quality control, assessment of authenticity and stability evaluation of ginseng flower buds. PMID:26270079

  18. The final cut: cell polarity meets cytokinesis at the bud neck in S. cerevisiae.

    Science.gov (United States)

    Juanes, Maria Angeles; Piatti, Simonetta

    2016-08-01

    Cell division is a fundamental but complex process that gives rise to two daughter cells. It includes an ordered set of events, altogether called "the cell cycle", that culminate with cytokinesis, the final stage of mitosis leading to the physical separation of the two daughter cells. Symmetric cell division equally partitions cellular components between the two daughter cells, which are therefore identical to one another and often share the same fate. In many cases, however, cell division is asymmetrical and generates two daughter cells that differ in specific protein inheritance, cell size, or developmental potential. The budding yeast Saccharomyces cerevisiae has proven to be an excellent system to investigate the molecular mechanisms governing asymmetric cell division and cytokinesis. Budding yeast is highly polarized during the cell cycle and divides asymmetrically, producing two cells with distinct sizes and fates. Many components of the machinery establishing cell polarization during budding are relocalized to the division site (i.e., the bud neck) for cytokinesis. In this review we recapitulate how budding yeast cells undergo polarized processes at the bud neck for cell division. PMID:27085703

  19. Glutamate may be an efferent transmitter that elicits inhibition in mouse taste buds.

    Directory of Open Access Journals (Sweden)

    Yijen A Huang

    Full Text Available Recent studies suggest that l-glutamate may be an efferent transmitter released from axons innervating taste buds. In this report, we determined the types of ionotropic synaptic glutamate receptors present on taste cells and that underlie this postulated efferent transmission. We also studied what effect glutamate exerts on taste bud function. We isolated mouse taste buds and taste cells, conducted functional imaging using Fura 2, and used cellular biosensors to monitor taste-evoked transmitter release. The findings show that a large fraction of Presynaptic (Type III taste bud cells (∼50% respond to 100 µM glutamate, NMDA, or kainic acid (KA with an increase in intracellular Ca(2+. In contrast, Receptor (Type II taste cells rarely (4% responded to 100 µM glutamate. At this concentration and with these compounds, these agonists activate glutamatergic synaptic receptors, not glutamate taste (umami receptors. Moreover, applying glutamate, NMDA, or KA caused taste buds to secrete 5-HT, a Presynaptic taste cell transmitter, but not ATP, a Receptor cell transmitter. Indeed, glutamate-evoked 5-HT release inhibited taste-evoked ATP secretion. The findings are consistent with a role for glutamate in taste buds as an inhibitory efferent transmitter that acts via ionotropic synaptic glutamate receptors.

  20. Programmed cell death during terminal bud senescence in a sympodial branching tree,Eucommia ulmoides

    Institute of Scientific and Technical Information of China (English)

    XU Wenjie; Kalima-N'Koma MWANGE; CUI Keming

    2004-01-01

    Eucommia ulmoides Oliv. is a typical sympodial branching tree. The apical bud of the branch ages and dies every year, replaced by the nearby axillary bud in the second year. Structural assays and a series of biochemical analyses were performed to analyze the senescence mechanism in the apical bud. It was revealed that most cells of the apical bud underwent the programmed cell death (PCD) during the senescence: the chromosomes were congregated and the nuclear contents were condensed, as shown by 4′,6-diamidino-2-phenylindole (DAPI) fluorescence. DNA fragmentation was detected during senescence using the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end in situ labeling (TUNEL) method, coincident with the appearance of a DNA ladder. Moreover, a 20 kD DNase related to fragmentation was found. PCD was initiated first in the young leaves, leaf primordia and peripheral zone cells, then in the central mother cells and initial layer cells in the apical meristem. The terminal buds remain in vegetative growth during senescence, in contrast to buds of many annual plants.

  1. Consequences of Repeated Defoliation on Belowground Bud Banks of Carex brevicuspis (Cyperaceae) in the Dongting Lake Wetlands, China

    Science.gov (United States)

    Chen, Xin-Sheng; Deng, Zheng-Miao; Xie, Yong-Hong; Li, Feng; Hou, Zhi-Yong; Wu, Chao

    2016-01-01

    Despite the predominant role of bud banks in the regeneration of clonal macrophyte populations, few studies have examined the way in which clonal macrophytes adjust the demographic features of bud banks to regulate population dynamics in response to defoliation in wetlands. We investigated the density and composition of bud banks under repeated defoliation in the wetland sedge Carex brevicuspis C. B. Clarke in the Dongting Lake wetlands, China. The density and biomass of rhizome buds and shoots did not decrease significantly in response to repeated defoliation over two consecutive years. The composition of bud banks, which consisted of long and short rhizome buds, also did not change significantly in response to repeated defoliation. Nevertheless, the ramet height and the shoot, root, and rhizome mass of C. brevicuspis declined significantly under repeated defoliation. Our findings suggest that bud banks are a conservative reproductive strategy that enables C. brevicuspis to tolerate a certain amount of defoliation. The maintenance of large bud banks after repeated defoliation may enable C. brevicuspis populations to regenerate and persist in disturbed habitats. However, bud bank density of C. brevicuspis might decline in the long term because the amount of carbon stored in rhizome buds and plants is reduced by frequent defoliation. PMID:27524993

  2. The cellular robustness by genetic redundancy in budding yeast.

    Directory of Open Access Journals (Sweden)

    Jingjing Li

    2010-11-01

    Full Text Available The frequent dispensability of duplicated genes in budding yeast is heralded as a hallmark of genetic robustness contributed by genetic redundancy. However, theoretical predictions suggest such backup by redundancy is evolutionarily unstable, and the extent of genetic robustness contributed from redundancy remains controversial. It is anticipated that, to achieve mutual buffering, the duplicated paralogs must at least share some functional overlap. However, counter-intuitively, several recent studies reported little functional redundancy between these buffering duplicates. The large yeast genetic interactions released recently allowed us to address these issues on a genome-wide scale. We herein characterized the synthetic genetic interactions for ∼500 pairs of yeast duplicated genes originated from either whole-genome duplication (WGD or small-scale duplication (SSD events. We established that functional redundancy between duplicates is a pre-requisite and thus is highly predictive of their backup capacity. This observation was particularly pronounced with the use of a newly introduced metric in scoring functional overlap between paralogs on the basis of gene ontology annotations. Even though mutual buffering was observed to be prevalent among duplicated genes, we showed that the observed backup capacity is largely an evolutionarily transient state. The loss of backup capacity generally follows a neutral mode, with the buffering strength decreasing in proportion to divergence time, and the vast majority of the paralogs have already lost their backup capacity. These observations validated previous theoretic predictions about instability of genetic redundancy. However, departing from the general neutral mode, intriguingly, our analysis revealed the presence of natural selection in stabilizing functional overlap between SSD pairs. These selected pairs, both WGD and SSD, tend to have decelerated functional evolution, have higher propensities of co

  3. Systems Level Modeling of the Cell Cycle Using Budding Yeast

    Directory of Open Access Journals (Sweden)

    D.R. Kim

    2007-01-01

    Full Text Available Proteins involved in the regulation of the cell cycle are highly conserved across all eukaryotes, and so a relatively simple eukaryote such as yeast can provide insight into a variety of cell cycle perturbations including those that occur in human cancer. To date, the budding yeast Saccharomyces cerevisiae has provided the largest amount of experimental and modeling data on the progression of the cell cycle, making it a logical choice for in-depth studies of this process. Moreover, the advent of methods for collection of high-throughput genome, transcriptome, and proteome data has provided a means to collect and precisely quantify simultaneous cell cycle gene transcript and protein levels, permitting modeling of the cell cycle on the systems level. With the appropriate mathematical framework and suffi cient and accurate data on cell cycle components, it should be possible to create a model of the cell cycle that not only effectively describes its operation, but can also predict responses to perturbations such as variation in protein levels and responses to external stimuli including targeted inhibition by drugs. In this review, we summarize existing data on the yeast cell cycle, proteomics technologies for quantifying cell cycle proteins, and the mathematical frameworks that can integrate this data into representative and effective models. Systems level modeling of the cell cycle will require the integration of high-quality data with the appropriate mathematical framework, which can currently be attained through the combination of dynamic modeling based on proteomics data and using yeast as a model organism.

  4. Differentiated dynamics of bud dormancy and growth in temperate fruit trees relating to bud phenology adaptation, the case of apple and almond trees

    Science.gov (United States)

    El Yaacoubi, Adnane; Malagi, Gustavo; Oukabli, Ahmed; Citadin, Idemir; Hafidi, Majida; Bonhomme, Marc; Legave, Jean-Michel

    2016-04-01

    Few studies have focused on the characterization of bud dormancy and growth dynamics for temperate fruit species in temperate and mild cropping areas, although this is an appropriate framework to anticipate phenology adaptation facing future warming contexts which would potentially combine chill declines and heat increases. To examine this issue, two experimental approaches and field observations were used for high- and low-chill apple cultivars in temperate climate of southern France and in mild climates of northern Morocco and southern Brazil. Low-chill almond cultivars offered an additional relevant plant material for comparison with apple in northern Morocco. Divergent patterns of dormancy and growth dynamics were clearly found in apple tree between southern France and southern Brazil. Divergences were less pronounced between France and Morocco. A global view outlined main differences in the dormancy chronology and intensity, the transition between endordormancy and ecodormancy and the duration of ecodormancy. A key role of bud rehydration in the transition period was shown. High-chill cultivars would be submitted in mild conditions to heterogeneous rehydration capacities linked to insufficient chill fulfillment and excessive forcing linked to high temperatures. This would favor bud competitions and consequently excessive flowering durations and weak flowering. Low chilling requirements in apple and almond would conversely confer biological capacities to tolerate superficial dormancy and abrupt transition from endordormancy to ecodormancy without important heterogeneous rehydration states within buds. It may also assume that low-chill cultivars can also tolerate high temperatures during ecodormancy as well as extended flowering durations.

  5. 5'-end sequences of budding yeast full-length cDNA clones and quality scores - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available Budding yeast cDNA sequencing project 5'-end sequences of budding yeast full-length cDNA clones and quality ...scores Data detail Data name 5'-end sequences of budding yeast full-length cDNA clones and quality scores De...from the budding yeast full-length cDNA library by the vector-capping method, the sequence quality score gen...s accession only. Sequence 5'-end sequence data of budding yeast full-length cDNA clones. FASTA format. Quality Phred's quality... Update History of This Database Site Policy | Contact Us 5'-end sequences of budding yeast full-length cDNA clones and quality

  6. High biological variability of plastids, photosynthetic pigments and pigment forms of leaf primordia in buds.

    Science.gov (United States)

    Solymosi, Katalin; Morandi, Dominique; Bóka, Károly; Böddi, Béla; Schoefs, Benoît

    2012-05-01

    To study the formation of the photosynthetic apparatus in nature, the carotenoid and chlorophyllous pigment compositions of differently developed leaf primordia in closed and opening buds of common ash (Fraxinus excelsior L.) and horse chestnut (Aesculus hippocastanum L.) as well as in closed buds of tree of heaven (Ailanthus altissima P. Mill.) were analyzed with HPLC. The native organization of the chlorophyllous pigments was studied using 77 K fluorescence spectroscopy, and plastid ultrastructure was investigated with electron microscopy. Complete etiolation, i.e., accumulation of protochlorophyllide, and absence of chlorophylls occurred in the innermost leaf primordia of common ash buds. The other leaf primordia were partially etiolated in the buds and contained protochlorophyllide (0.5-1 μg g(-1) fresh mass), chlorophyllides (0.2-27 μg g(-1) fresh mass) and chlorophylls (0.9-643 μg g(-1) fresh mass). Etio-chloroplasts with prolamellar bodies and either regular or only low grana were found in leaves having high or low amounts of chlorophyll a and b, respectively. After bud break, etioplast-chloroplast conversion proceeded and the pigment contents increased in the leaves, similarly to the greening processes observed in illuminated etiolated seedlings under laboratory conditions. The pigment contents and the ratio of the different spectral forms had a high biological variability that could be attributed to (i) various light conditions due to light filtering in the buds resulting in differently etiolated leaf primordia, (ii) to differences in the light-exposed and inner regions of the same primordia in opening buds due to various leaf folding, and (iii) to tissue-specific slight variations of plastid ultrastructure. PMID:22160501

  7. Local adaptations and climate change: converging sensitivity of bud break in black spruce provenances

    Science.gov (United States)

    Rossi, Sergio

    2015-07-01

    Species with transcontinental distribution or spread over wide geographical regions develop populations with growth traits genetically adapted to the local climate. The aim of this study was to investigate the ecotypic sensitivity of bud break, a strong adaptive trait, to a changing environment. Six phenological phases of bud break were monitored daily on black spruce [ Picea mariana (Mill.) BSP] seedlings submitted to different temperatures (12, 16 and 20 °C) and photoperiods (14, 18 and 22 h). Six provenances were tested in growth chambers, produced from seeds collected along the whole latitudinal range of the closed boreal forest in Quebec, Canada. Bud break lasted 13.3 days on average and occurred earlier in seedlings from colder sites. The annual temperature of the sites suitably tracked the clinal variation among ecotypes, providing a clear biological explanation for the environmental signal driving the adaptive divergence of populations to the local climate. Increasing temperature induced an earlier bud break according to a non-linear pattern with greater advancements observed between 12 and 16 °C. Photoperiod was significant, but sensitivity analysis indicated that its effect on bud break was marginal with respect to temperature. No interaction of provenance × treatment was observed, demonstrating an ecotypic convergence of the responses to both factors. Changes in the growing conditions could substantially modify the synchronization between bud phenology and climate, thus exposing the developing meristems of black spruce to frost damage. However, similar advancements of bud break could be expected in the different ecotypes subjected to warmer temperatures or longer day lengths.

  8. Reactivation from latency displays HIV particle budding at plasma membrane, accompanying CD44 upregulation and recruitment

    Directory of Open Access Journals (Sweden)

    Sano Kouichi

    2009-07-01

    Full Text Available Abstract Background It has been accepted that HIV buds from the cell surface in T lymphocytes, whereas in macrophages it buds into intracellular endosomes. Recent studies, on the other hand, suggest that HIV preferentially buds from the cell surface even in monocytic cells. However, most studies are based on observations in acutely infected cells and little is known about HIV budding concomitant with reactivation from latency. Such studies would provide a better understanding of a reservoir for HIV. Results We observed HIV budding in latently infected T lymphocytic and monocytic cell lines following TNF-α stimulation and examined the upregulation of host factors that may be involved in particle production. Electron microscopy analysis revealed that reactivation of latently infected J1.1 cells (latently infected Jurkat cells with HIV-1 and U1 cells (latently infected U937 cells with HIV-1 displayed HIV particle budding predominantly at the plasma membrane, a morphology that is similar to particle budding in acutely infected Jurkat and U937 cells. When mRNA expression levels were quantified by qRT-PCR, we found that particle production from reactivated J1.1 and U1 cells was accompanied by CD44 upregulation. This upregulation was similarly observed when Jurkat and U937 cells were acutely infected with HIV-1 but not when just stimulated with TNF-α, suggesting that CD44 upregulation was linked with HIV production but not with cell stimulation. The molecules in endocytic pathways such as CD63 and HRS were also upregulated when U1 cells were reactivated and U937 cells were acutely infected with HIV-1. Confocal microscopy revealed that these upregulated host molecules were recruited to and accumulated at the sites where mature particles were formed at the plasma membrane. Conclusion Our study indicates that HIV particles are budded at the plasma membrane upon reactivation from latency, a morphology that is similar to particle budding in acute

  9. Relationship between sensitivity to ultraviolet light and budding in yeast cells of different culture ages

    International Nuclear Information System (INIS)

    Subpopulations of yeast cells, consisting of cells of different sizes and different percentages of budding cells, were prepared by centrifugation through sucrose solutions with linear density gradients of cultures at different phases of the growth cycle. Ultraviolet survival of these cells was determined by colony counting, and the survival rate was compared with the cells' respiratory rates. Individual budding cells and interdivisional cells, and also mother cells and daughter cells derived from irradiated budding cells, were isolated by the micromanipulation technique. The number of divisions in each cell was measured during a 21-hr incubation period immediately after irradiation. In the population in the logarithmic phase consisting of homogeneous cells of middle size, no difference in uv sensitivity was observed between mother cells and daughter cells, irrespective of mutual adhesion. Budding cell resistance was observed in the population in the transitional phase; this was due to the lesser uv sensitivity of daughter cells in the fresh medium. In the stationary phase, daughter cells were rather more sensitive than mother cells or interdivisional cells, so there was little difference in uv sensitivity between budding cells and interdivisional cells

  10. Study of budding yeast colony formation and its characterizations by using circular granular cell

    Science.gov (United States)

    Aprianti, D.; Haryanto, F.; Purqon, A.; Khotimah, S. N.; Viridi, S.

    2016-03-01

    Budding yeast can exhibit colony formation in solid substrate. The colony of pathogenic budding yeast can colonize various surfaces of the human body and medical devices. Furthermore, it can form biofilm that resists drug effective therapy. The formation of the colony is affected by the interaction between cells and with its growth media. The cell budding pattern holds an important role in colony expansion. To study this colony growth, the molecular dynamic method was chosen to simulate the interaction between budding yeast cells. Every cell was modelled by circular granular cells, which can grow and produce buds. Cohesion force, contact force, and Stokes force govern this model to mimic the interaction between cells and with the growth substrate. Characterization was determined by the maximum (L max) and minimum (L min) distances between two cells within the colony and whether two lines that connect the two cells in the maximum and minimum distances intersect each other. Therefore, it can be recognized the colony shape in circular, oval, and irregular shapes. Simulation resulted that colony formation are mostly in oval shape with little branch. It also shows that greater cohesion strength obtains more compact colony formation.

  11. ALG-2 attenuates COPII budding in vitro and stabilizes the Sec23/Sec31A complex.

    Directory of Open Access Journals (Sweden)

    Jonas M la Cour

    Full Text Available Coated vesicles mediate the traffic of secretory and membrane cargo proteins from the endoplasmic reticulum (ER to the Golgi apparatus. The coat protein complex (COPII involved in vesicle budding is constituted by a GTPase, Sar1, the inner coat components of Sec23/Sec24 and the components of the outer coat Sec13/Sec31A. The Ca(2+-binding protein ALG-2 was recently identified as a Sec31A binding partner and a possible link to Ca(2+ regulation of COPII vesicle budding. Here we show that ALG-2/Ca(2+ is capable of attenuating vesicle budding in vitro through interaction with an ALG-2 binding domain in the proline rich region of Sec31A. Binding of ALG-2 to Sec31A and inhibition of COPII vesicle budding is furthermore dependent on an intact Ca(2+-binding site at EF-hand 1 of ALG-2. ALG-2 increased recruitment of COPII proteins Sec23/24 and Sec13/31A to artificial liposomes and was capable of mediating binding of Sec13/31A to Sec23. These results introduce a regulatory role for ALG-2/Ca(2+ in COPII tethering and vesicle budding.

  12. A2BR adenosine receptor modulates sweet taste in circumvallate taste buds.

    Directory of Open Access Journals (Sweden)

    Shinji Kataoka

    Full Text Available In response to taste stimulation, taste buds release ATP, which activates ionotropic ATP receptors (P2X2/P2X3 on taste nerves as well as metabotropic (P2Y purinergic receptors on taste bud cells. The action of the extracellular ATP is terminated by ectonucleotidases, ultimately generating adenosine, which itself can activate one or more G-protein coupled adenosine receptors: A1, A2A, A2B, and A3. Here we investigated the expression of adenosine receptors in mouse taste buds at both the nucleotide and protein expression levels. Of the adenosine receptors, only A2B receptor (A2BR is expressed specifically in taste epithelia. Further, A2BR is expressed abundantly only in a subset of taste bud cells of posterior (circumvallate, foliate, but not anterior (fungiform, palate taste fields in mice. Analysis of double-labeled tissue indicates that A2BR occurs on Type II taste bud cells that also express Gα14, which is present only in sweet-sensitive taste cells of the foliate and circumvallate papillae. Glossopharyngeal nerve recordings from A2BR knockout mice show significantly reduced responses to both sucrose and synthetic sweeteners, but normal responses to tastants representing other qualities. Thus, our study identified a novel regulator of sweet taste, the A2BR, which functions to potentiate sweet responses in posterior lingual taste fields.

  13. Study of disbudding goat kids following injection of clove oil essence in horn bud region.

    Science.gov (United States)

    Molaei, Mohammad Mahdi; Mostafavi, Ali; Kheirandish, Reza; Azari, Omid; Shaddel, Mohsen

    2015-01-01

    This study was performed to evaluate the efficacy of injection of essential oil of Eugenia caryophyllata in the kid horn buds, as a new chemical technique for disbudding. Five-day-old healthy goat kids from both sexes (n = 16) were divided randomly into 4 equal groups. In groups 1, 2 and 3, 0.2 mL of clove essence and in group 4 (control) 0.2 mL of normal saline was injected into the left horn bud of goat kids. Right horn bud in all kids was considered to ensure that they are horned. During the study, the rate of horn growth were evaluated in determined time intervals between groups 1 and 4. Tissue samples were taken from right and left horn bud in groups 2 and 3, at five and ten days after clove essence injection, for microscopic study. The results of the study showed that the clove essence stopped horn growth, whereas there was no significant difference in horn growth rate between left and right horns after injection of normal saline, in group 4. Histopathological study showed that injection of clove essence caused complete necrosis of epidermis and underlying dermis with collagenolysis in horn bud tissues, 5 days after injection and then progress in healing process was observed after 10 days. According to the results of this study, it can be concluded that the injection of clove essence is an effective method to stop horn growth without any undesirable effects on clinical parameters in goat kids. PMID:25992247

  14. Relation Between Endodormancy Induction and Changes in Two Main Electron Transport Pathways of Nectarine Buds

    Institute of Scientific and Technical Information of China (English)

    YU Qin; GAO Dong-sheng; XU Xiao-ming; LI Jin; XU Chen-shan

    2008-01-01

    Operation regulations of two main electron transport pathways in nectarine (Prunus persica var. nectariana cv. Shuguang) buds during endodormancy induction were studied to understand possible roles which two main electron transport pathways played in the buds of deciduous fruit trees during endodormancy induction. Respiratory inhibitors (KCN and SHAM) were used to investigate total respiration rate (Vt), the development and operation of the alternative pathway and partitioning of electrons between the cytochrome and alternative pathways in nectarine buds during endodormancy induction. Results indicated that changes of V, in flower and leaf buds showed single and double hump-shaped curves, respectively. In endodormancy induction, the capacity (V^,,) and activity (pValt) of the alternative pathway rapidly increased, but changes of them had different patterns during the entire measuration. At the same time, changes of engagements of the alternative (pValt/Vt) and cytochrome pathway (p'Vcy/Vt) were opposite, and p'Vcyt/Vt was always further higher than pValt/Vt during the entire measuration. All these results indicated that the development and operation of the alternative pathway played important roles in endodormancy induction, but the cytochrome pathway was the main pathway for mitochondrial electron transport in buds during endodormancy induction.

  15. Respiratory Response of Dormant Nectarine Vegetative Buds to High Temperature Stress

    Institute of Scientific and Technical Information of China (English)

    TAN Yue; LI Ling; LENG Chuan-yuan; LI Dong-mei; CHEN Xiu-de; GAO Dong-sheng

    2013-01-01

    High temperature stress (HT) is efficient in breaking endo-dormancy of perennial trees. The effects of HT (50°C) on the respiration of dormant nectarine (Prunus persica var. nectariana cv. Shuguang) vegetative buds were evaluated in the research. We found that bud respiration was transiently inhibited by HT and the pentose phosphate pathway (PPP) and the cytochrome C pathway (CYT) were significantly affected. On the substrate level, PPP was activated in the HT-treated buds compared with the control group. However, the activation did mot occur until hours after HT treatment. The tricarboxylic acid cycle (TCA) in both the HT-treated buds and in the control group proceeded at a low level most of the time compared with total respiration. On the electron transfer level, CYT was transiently inhibited by HT but became significantly active in the later stage. CYT operation in the control group exhibited an attenuation process. The alternative pathway (ALT) fluctuated both in the HT-treated samples and in the control. The results suggest that the temporary CYT inhibition and the following PPP activation may be involved in HT-induced bud dormancy release and budburst mechanisms.

  16. Ferrite-Cored Solenoidal Induction Coil Sensor for BUD (MM-1667)

    Energy Technology Data Exchange (ETDEWEB)

    Morrison, F.; Becker, A.; Conti, U.; Gasperikova, E.

    2011-06-15

    We have designed and lab tested a new ferrite cored induction coil sensor for measuring the secondary fields from metallic UXO with the BUD system. The objective was to replace the 5-inch diameter air-cored coils in the BUD system with smaller sensors that would allow the placement of multiple sensors in the smaller package of the new BUD hand-held system. A ferrite-cored solenoidal coil of length L can easily be made to have sensitivity and noise level roughly the same as an air-cored coil of a diameter on the same order as L. A ferrite-cored solenoidal coil can easily have a feedback configuration to achieve critical damping. The feedback configuration leads to a very stable response. Feedback ferrite-cored solenoidal coils show very little interaction as long as they are separated by one half their length.

  17. Multiple shoot-bud formation and plantlet regeneration on Castanea sativa Mill. seeds in culture.

    Science.gov (United States)

    Rodríguez, R

    1982-06-01

    Primordial initiation and development of shoot-buds has been accomplished by using shoots derived from chestnut (Castanea sativa Mill) seedlings cultured with added 6-benzylaminopurine (BAP). Germination of chestnut seeds in the presence of BAP (4 - 40 μM) stimulated varying numbers of shoot-buds in those areas of the main axis that were favorably altered. When excised single shoots from these treated seeds were subcultured on a fresh medium containing BAP (4 - 40 μM) continual shoot production was observed. Bud growth and shoot elongation were stimulated by transferring cultures to a reduced concentration of BAP (2 μM) plus indole-3-butyric acid (IBA 0.4 μM). Plant regeneration occurred in the presence of IBA (0.8 μM) after a preconditioning treatment in which naphthaleneacetic acid (NAA 50 μM) and kinetin (k 2 μM) were applied to the tissue culture shoots for 7 days in light.

  18. Progress and renewal in gustation: new insights into taste bud development.

    Science.gov (United States)

    Barlow, Linda A

    2015-11-01

    The sense of taste, or gustation, is mediated by taste buds, which are housed in specialized taste papillae found in a stereotyped pattern on the surface of the tongue. Each bud, regardless of its location, is a collection of ∼100 cells that belong to at least five different functional classes, which transduce sweet, bitter, salt, sour and umami (the taste of glutamate) signals. Taste receptor cells harbor functional similarities to neurons but, like epithelial cells, are rapidly and continuously renewed throughout adult life. Here, I review recent advances in our understanding of how the pattern of taste buds is established in embryos and discuss the cellular and molecular mechanisms governing taste cell turnover. I also highlight how these findings aid our understanding of how and why many cancer therapies result in taste dysfunction.

  19. Greenhouse irrigation water depths in relation to rose stem and bud qualities

    Directory of Open Access Journals (Sweden)

    Folegatti Marcos Vinícius

    2001-01-01

    Full Text Available The cultivation of roses occupies a special place in the flower production of Brazil, the concern with the quality of the buds being intimately related with the appropriate supply of water and nutrients to the plant. With the objective of evaluating stem and bud quality the rose variety 'Osiana' was cultivated in a greenhouse using different irrigation water depths based on fractions of pan evaporation (0.25, 0.50, 0.75, 1.00 and 1.25. The experimental design consisted of total randomized blocks with five replications and five treatments. There is a linear tendency of increasing the length and diameter of the stems and the length and diameter of the buds with increasing irrigation water depths.

  20. Progress and renewal in gustation: new insights into taste bud development.

    Science.gov (United States)

    Barlow, Linda A

    2015-11-01

    The sense of taste, or gustation, is mediated by taste buds, which are housed in specialized taste papillae found in a stereotyped pattern on the surface of the tongue. Each bud, regardless of its location, is a collection of ∼100 cells that belong to at least five different functional classes, which transduce sweet, bitter, salt, sour and umami (the taste of glutamate) signals. Taste receptor cells harbor functional similarities to neurons but, like epithelial cells, are rapidly and continuously renewed throughout adult life. Here, I review recent advances in our understanding of how the pattern of taste buds is established in embryos and discuss the cellular and molecular mechanisms governing taste cell turnover. I also highlight how these findings aid our understanding of how and why many cancer therapies result in taste dysfunction. PMID:26534983

  1. Bud flush phenology and nursery carryover effect of paper birch provenances

    Directory of Open Access Journals (Sweden)

    Dhar A

    2015-12-01

    Full Text Available Paper birch (Betula papyrifera Marsh is an ecologically valuable species with a broad geographic distribution across the North America. Its diversity, versatility and enduring nature make it an ideal candidate for a selective breeding program in this region. However, an understanding of the genecology of this species is fundamental to deploy it successfully. Ten paper birch provenances were collected from British Columbia (BC, Canada and northern Idaho (USA along elevational transects to determine whether observed bud flush phenology was due to genetics and /or environmental variation or their interaction. Seedlings were grown at three different nurseries: University of Idaho (46°44’N, Landing (50°17’N and Little Forestry (54°00’N and planted in a randomized single tree interlocking block design in three common gardens at Sandpoint, ID (48°13’N, Skimikin, BC (50°45’N and Red Rock, BC (53°45’N. Results indicate that variation in the timing of bud flush is a complex interaction among local genetic characteristics and environmental conditions of the growing site. Birch bud flush followed a general geographic trend where provenances at the southern common garden (Sandpoint required less time (Day of Year, DoY and fewer growing degree days (GDD compared to central (Skimikin and northern (Red Rock common gardens. Although there were significant differences in the timing of bud flush among provenances along an elevational gradient, none of the regions showed the expected linear elevational cline, trends were inconsistent. Further, birch bud flush was significantly influenced by nursery displacement effects in the initial year of establishment but disappeared within three years. These results provide an opportunity to characterize bud flush phenology of paper birch and would be useful for improving operational paper birch seed transfer programs in BC.

  2. Lamin Mutations Accelerate Aging via Defective Export of Mitochondrial mRNAs through Nuclear Envelope Budding.

    Science.gov (United States)

    Li, Yihang; Hassinger, Linda; Thomson, Travis; Ding, Baojin; Ashley, James; Hassinger, William; Budnik, Vivian

    2016-08-01

    Defective RNA metabolism and transport are implicated in aging and degeneration [1, 2], but the underlying mechanisms remain poorly understood. A prevalent feature of aging is mitochondrial deterioration [3]. Here, we link a novel mechanism for RNA export through nuclear envelope (NE) budding [4, 5] that requires A-type lamin, an inner nuclear membrane-associated protein, to accelerated aging observed in Drosophila LaminC (LamC) mutations. These LamC mutations were modeled after A-lamin (LMNA) mutations causing progeroid syndromes (PSs) in humans. We identified mitochondrial assembly regulatory factor (Marf), a mitochondrial fusion factor (mitofusin), as well as other transcripts required for mitochondrial integrity and function, in a screen for RNAs that exit the nucleus through NE budding. PS-modeled LamC mutations induced premature aging in adult flight muscles, including decreased levels of specific mitochondrial protein transcripts (RNA) and progressive mitochondrial degradation. PS-modeled LamC mutations also induced the accelerated appearance of other phenotypes associated with aging, including a progressive accumulation of polyubiquitin aggregates [6, 7] and myofibril disorganization [8, 9]. Consistent with these observations, the mutants had progressive jumping and flight defects. Downregulating marf alone induced the above aging defects. Nevertheless, restoring marf was insufficient for rescuing the aging phenotypes in PS-modeled LamC mutations, as other mitochondrial RNAs are affected by inhibition of NE budding. Analysis of NE budding in dominant and recessive PS-modeled LamC mutations suggests a mechanism by which abnormal lamina organization prevents the egress of these RNAs via NE budding. These studies connect defects in RNA export through NE budding to progressive loss of mitochondrial integrity and premature aging. PMID:27451905

  3. 6-benzyladenine metabolism during reinvigoration of mature Pinus radiata buds in vitro.

    Science.gov (United States)

    Zhang, Huaibi; Horgan, Kathryn J; Reynolds, Paul H S; Jameson, Paula E

    2010-04-01

    Maturation or phase change is a serious challenge in the deployment of superior trees of Pinus radiata D. Don because of the difficulties associated with propagation of cuttings from mature trees. We used an in vitro system to study 6-benzyladenine (BA)-induced reinvigoration of the fascicle meristems of mature buds during in vitro culture. Anatomical examinations revealed that BA inhibited the development of secondary needle primordia and 'rejuvenated' the fascicle meristems of the mature bud to produce primary needles, which are characteristic of the juvenile phase in P. radiata. Without BA supplement in the culture media, fascicle primordia continued developing secondary needles and quiescent fascicle meristems. BA metabolite analysis showed that the novel cytokinin pathway reported previously in P. radiata (H. Zhang, K.J. Horgan, P.H.S. Reynolds, G.E. Norris and P.E. Jameson. 2001. Novel cytokinins: The predominant forms in mature buds of Pinus radiata. Physiol. Plant. 112: 127-134) was mirrored in vitro, with BA converted into a variety of metabolites including 6-benzylamino-9-glucopyranosylribosyl-purine and its novel phosphorylated form, 6-benzylamino-9-glucopyranosylribosyl-purine. The culture of mature buds in the presence of BA caused a reduction in the level of endogenous cytokinins, suggesting a direct action of BA itself. Similar correlations are noted between levels of certain metabolites and the maturation status of buds from field-grown trees and buds in culture, indicating that this in vitro system may be a good model for studying the processes of maturation and reinvigoration. PMID:20144924

  4. Dormancy release and chilling requirement of buds of latitudinal ecotypes of Betula pendula and B. pubescens.

    Science.gov (United States)

    Myking, T; Heide, O M

    1995-11-01

    Bud burst and dormancy release of latitudinal ecotypes of Betula pendula Roth and B. pubescens Ehrh. from Denmark ( approximately 56 degrees N), mid-Norway ( approximately 64 degrees N) and northern Norway ( approximately 69 degrees N) were studied in controlled environments. Dormant seedlings were chilled at 0, 5 or 10 degrees C from October 4 onward and then, at monthly intervals from mid-November to February, batches of seedlings were held at 15 degrees C in an 8-h (SD) or 24-h (LD) photoperiod to permit flushing. A decline in days to bud burst occurred with increasing chilling time in all ecotypes. In November, after 44 chilling days, time to bud burst was least in plants chilled at 0 and 5 degrees C. The difference diminished with increasing chilling time, and in February, after 136 chilling days, bud burst was earliest in plants chilled at 10 degrees C. Long photoperiods during flushing significantly reduced thermal time after short chilling periods (44 and 74 days), but had no effect when the chilling requirement was fully met after 105 or more chilling days. No significant difference in these responses was found between the two species. In both species, chilling requirement decreased significantly with increasing latitude of origin. Bud burst was normal in seedlings overwintered at 12 degrees C, but was erratic and delayed in seedlings overwintered at 15 and especially at 21 degrees C, indicating that the critical overwintering temperature is between 12 and 15 degrees C. We conclude that there is little risk of a chilling deficit in birch under Scandinavian winter conditions even with a climatic warming of 7-8 degrees C. The likely effects of a climatic warming include earlier bud burst, a longer growing season and increased risk of spring frost injury, especially in high latitude ecotypes. PMID:14965987

  5. ESCRT-independent budding of HIV-1 gag virus-like particles from Saccharomyces cerevisiae spheroplasts.

    Directory of Open Access Journals (Sweden)

    Andrew P Norgan

    Full Text Available Heterologous expression of HIV-1 Gag in a variety of host cells results in its packaging into virus-like particles (VLPs that are subsequently released into the extracellular milieu. This phenomenon represents a useful tool for probing cellular factors required for viral budding and has contributed to the discovery of roles for ubiquitin ligases and the endosomal sorting complexes required for transport (ESCRTs in viral budding. These factors are highly conserved throughout eukaryotes and have been studied extensively in the yeast Saccharomyces cerevisiae, a model eukaryote previously utilized as a host for the production of VLPs. We used heterologous expression of HIV Gag in yeast spheroplasts to examine the role of ESCRTs and associated factors (Rsp5, a HECT ubiquitin ligase of the Nedd4 family; Bro1, a homolog of Alix; and Vps4, the AAA-ATPase required for ESCRT function in all contexts/organisms investigated in the generation of VLPs. Our data reveal: 1 characterized Gag-ESCRT interaction motifs (late domains are not required for VLP budding, 2 loss of function alleles of the essential HECT ubiquitin ligase Rsp5 do not display defects in VLP formation, and 3 ESCRT function is not required for VLP formation from spheroplasts. These results suggest that the egress of HIV Gag from yeast cells is distinct from the most commonly described mode of exit from mammalian cells, instead mimicking ESCRT-independent VLP formation observed in a subset of mammalian cells. As such, budding of Gag from yeast cells appears to represent ESCRT-independent budding relevant to viral replication in at least some situations. Thus the myriad of genetic and biochemical tools available in the yeast system may be of utility in the study of this aspect of viral budding.

  6. Pickling process of capers (Capparis spp.) flower buds

    OpenAIRE

    Özcan, Musa; Akgül, Attila

    1999-01-01

    Middle sized (8 < x < 13 mm) buds of Capparis spinosa var. spinosa and C. ovata var. canescens from June in brines containing 5,10,15 and 20% salt and from August in brines of 15% salt, and three different size (x < 8 mm, 8 < x < 13 mm, x > 13 mm) buds of C. . ovata var. canescens from June in brines of 15% salt were pickled for two months fermentation. Some chem...

  7. Project BudBurst - Meeting the Needs of Climate Change Educators and Scientists

    Science.gov (United States)

    Henderson, S.

    2015-12-01

    It is challenging for many to get a sense of what climate change means as long periods of time are involved - like decades - which can be difficult to grasp. However, there are a number of citizen science based projects, including NEON's Project BudBurst, that provide the opportunity for both learning about climate change and advancing scientific knowledge. In this presentation, we will share lessons learned from Project BudBurst. Project BudBurst is a national citizen science initiative designed to engage the public in observations of phenological (plant life cycle) events and to increase climate literacy. Project BudBurst is important from an educational perspective, but also because it enables scientists to broaden the geographic and temporal scale of their observations. The goals of Project BudBurst are to 1) increase awareness of phenology as an area of scientific study; 2) Increase awareness of the impacts of changing climates on plants at a continental-scale; and 3) increase science literacy by engaging participants in the scientific process. It was important to better understand if and how Project BudBurst is meeting its goals. Specifically, does participation by non-experts advance scientific knowledge? Does participation advance educational goals and outcomes? Is participation an effective approach to advance/enhance science education in both formal and informal settings? Critical examination of Project BudBurst supports advancement of scientific knowledge and realization of educational objectives. Citizen science collected observations and measurements are being used by scientists as evidenced by the increase of such data in scientific publication. In addition, we found that there is a significant increase in educators utilizing citizen science as part of their instruction. Part of this increase is due to the resources and professional development materials available to educators. Working with partners also demonstrated that the needs of both science and

  8. Chemical composition and content of essential oil from the bud of cultivated Turkish clove

    OpenAIRE

    Kollmannsberger, H.; Nitz, S.; Ertaş, M.; Alma, M. H.

    2007-01-01

    In this study, clove bud oil, which was cultivated in the Mediterranean region of Turkey, was provided from a private essential oil company in Turkey. Essential oil from clove (Syzygium aromaticum L.) was obtained from steam-distillation method, and its chemical composition was analyzed by GC and GC-MS. The results showed that the essential oils mainly contained about 87.00% eugenol, 8.01% eugenyl acetate and 3.56% β-Caryophyllene. The chemical composition of the Turkish clove bud oil was com...

  9. Download - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available Budding yeast cDNA sequencing project Download First of all, please read the license of this database. Data ...names and data descriptions are about the downloadable data in this page. They might not correspond to the c...f the data. # Data name File Simple search and download 1 README README_e.html - 2 5'-end sequences of buddi...ng yeast full-length cDNA clones and quality scores yeast_seq_qual.zip (59.9MB) Simple search and download 3...Downlaod via FTP Joomla SEF URLs by Artio About This Database Database Description Download License Update H

  10. Chemical Genetics: Budding Yeast as a Platform for Drug Discovery and Mapping of Genetic Pathways

    Directory of Open Access Journals (Sweden)

    Jorrit M. Enserink

    2012-08-01

    Full Text Available The budding yeast Saccharomyces cerevisiae is a widely used model organism, and yeast genetic methods are powerful tools for discovery of novel functions of genes. Recent advancements in chemical-genetics and chemical-genomics have opened new avenues for development of clinically relevant drug treatments. Systematic mapping of genetic networks by high-throughput chemical-genetic screens have given extensive insight in connections between genetic pathways. Here, I review some of the recent developments in chemical-genetic techniques in budding yeast.

  11. Budding yeast PAK kinases regulate mitotic exit by two different mechanisms

    OpenAIRE

    Chiroli, Elena; Fraschini, Roberta; Beretta, Alessia; Tonelli, Mariagrazia; Lucchini, Giovanna; Piatti, Simonetta

    2003-01-01

    We report the characterization of the dominant-negative CLA4t allele of the budding yeast CLA4 gene, encoding a member of the p21-activated kinase (PAK) family of protein kinases, which, together with its homologue STE20, plays an essential role in promoting budding and cytokinesis. Overproduction of the Cla4t protein likely inhibits both endogenous Cla4 and Ste20 and causes a delay in the onset of anaphase that correlates with inactivation of Cdc20/anaphase-promoting complex (APC)–dependent ...

  12. Chemical composition and content of essential oil from the bud of cultivated Turkish clove

    Directory of Open Access Journals (Sweden)

    Kollmannsberger, H.

    2007-05-01

    Full Text Available In this study, clove bud oil, which was cultivated in the Mediterranean region of Turkey, was provided from a private essential oil company in Turkey. Essential oil from clove (Syzygium aromaticum L. was obtained from steam-distillation method, and its chemical composition was analyzed by GC and GC-MS. The results showed that the essential oils mainly contained about 87.00% eugenol, 8.01% eugenyl acetate and 3.56% β-Caryophyllene. The chemical composition of the Turkish clove bud oil was comparable to those of trees naturally grown in their native regions.

  13. Epidemiology and effective control of Altenaria altenata, causal agent of dead (dormant) flower bud disease of pear

    NARCIS (Netherlands)

    Wenneker, M.; Joosten, N.N.; Anbergen, R.H.N.; Vink, P.; Bruggen, van A.S.

    2011-01-01

    Dead flower buds are a common phenomenon in pear culture in The Netherlands, Belgium and Mediterranean countries. Disease cases are also reported from South America. The disease is characterized by a partial or complete necrosis of flower buds during tree dormancy. The disease progresses during wint

  14. Culture Media Optimization through Response Surface Methodology for in vitro Shoot Bud Development of Solanum melongena L. for Micropropagation

    Directory of Open Access Journals (Sweden)

    Sila Bhattacharya

    2011-11-01

    Full Text Available Response surface methodology was used for the optimization of shoot bud response and shoot bud induction in leaf explants of Solanum melongena cultivar Arka Shirish. Three independent variables were evaluated for shoot bud response and shoot bud induction. The variables include the concentrations of nitrogen (N2, sucrose and growth regulator thidiazuron (TDZ. The shoot bud response for cultured explant was optimized at 4.34 g/l of total nitrogen, 2.65% of sucrose and 0.67 mg/l of TDZ with 95% response. The optimum medium conditions for shoot bud induction was found to be Murashige and Skoog (MS basal medium supplemented with 4.02 g/l of total nitrogen, 2.36% of sucrose and 1.0 mg/l of TDZ with 10 number of bud per explant. The shoot buds so formed were elongated in 0.5 mg/l 2,3,5-Triiodobenzoic acid (TIBA and 0.1 mg/l Gibberellic acid (GA3. The elongated shoots were rooted in MS with 1 mg/l Indole-3-butyric acid (IBA. The rooted plants were transferred to pots with farmyard manure upon hardening. This study has validation value for optimization of micropropagation protocol and is further useful in genetic transformation studies for Solanum melongena variety Arka Shirish to maximize regenerative response for automation.

  15. The Role of Leaves in Photocontrol of Flower Bud Abscission in Hibiscus rosa-sinensis L. 'Nairobi'

    NARCIS (Netherlands)

    Meeteren, van U.; Gelder, van A.

    2000-01-01

    When compared with exposure to darkness, exposing Hibiscus rosa-sinensis L. 'Nairobi' plants to red light (635 to 685 nm, 2.9 μmol?m-2?s-1) delayed flower bud abscission, while exposure to far-red light (705 to 755 nm, μmol?m-2?s-1) accelerated this process. Flower bud abscission in response to ligh

  16. Neurturin-GFRalpha2 signaling controls liver bud migration along the ductus venosus in the chick embryo.

    Science.gov (United States)

    Tatsumi, Norifumi; Miki, Rika; Katsu, Kenjiro; Yokouchi, Yuji

    2007-07-01

    During chick liver development, the liver bud arises from the foregut, invaginates into the septum transversum, and elongates along and envelops the ductus venosus. However, the mechanism of liver bud migration is only poorly understood. Here, we demonstrate that a GDNF family ligand involved in neuronal outgrowth and migration, neurturin (NRTN), and its receptor, GFRalpha2, are essential for liver bud migration. In the chick embryo, we found that GFRalpha2 was expressed in the liver bud and that NRTN was expressed in the endothelial cells of the ductus venosus. Inhibition of GFRalpha2 signaling suppressed liver bud elongation along the ductus venous without affecting cell proliferation and apoptosis. Moreover, ectopic expression of NRTN perturbed the directional migration along the ductus venosus, leading to splitting or ectopic branching of the liver. We showed that liver buds selectively migrated toward an NRTN-soaked bead in vitro. These data represent a new model for liver bud migration: NRTN secreted from endothelial cells functions as a chemoattractant to direct the migration of the GFRalpha2-expressing liver bud in early liver development.

  17. Dehydration and osmotic adjustment in apple stem tissue during winter as it relates to the frost resistance of buds.

    Science.gov (United States)

    Pramsohler, Manuel; Neuner, Gilbert

    2013-08-01

    In deciduous trees, measurement of stem water potential can be difficult during the leafless period in winter. By using thermocouple psychrometry, osmotic water potentials (Ψo; actual Ψo: Ψo(act); Ψo at full saturation: Ψo(sat)) of expressed sap of bark and bud tissue were measured in order to test if the severity of winter desiccation in apple stems could be sufficiently assessed with Ψo. Water potentials were related to frost resistance and freezing behaviour of buds. The determination of Ψo reliably allowed winter desiccation and osmotic adjustments in apple stem tissue to be assessed. In winter in bark tissue, a pronounced decrease in Ψo(act) and Ψo(sat) was found. Decreased Ψo(sat) indicates active osmotic adjustment in the bark as observed earlier in the leaves of evergreen woody plants. In terminal bud meristems, no significant osmotic adjustments occurred and dehydration during winter was much less. Osmotic water potentials, Ψo(act) and Ψo(sat), of bud tissue were always less negative than in the bark. To prevent water movement and dehydration of the bud tissue via this osmotic gradient, it must be compensated for either by a sufficiently high turgor pressure (Ψp) in bark tissue or by the isolation of the bud tissue from the bark during midwinter. During freezing of apple buds, freeze dehydration and extra-organ freezing could be demonstrated by significantly reduced Ψo(act) values of bud meristems that had been excised in the frozen state. Infrared video thermography was used to monitor freezing patterns in apple twigs. During extracellular freezing of intact and longitudinally dissected stems, infrared differential thermal analysis (IDTA) images showed that the bud meristem remains ice free. Even if cooled to temperatures below the frost-killing temperature, no freezing event could be detected in bud meristems during winter. In contrast, after bud break, terminal buds showed a second freezing at the frost-killing temperature that indicates

  18. The yeast prefoldin-like URI-orthologue Bud27 associates with the RSC nucleosome remodeler and modulates transcription.

    Science.gov (United States)

    Mirón-García, María Carmen; Garrido-Godino, Ana Isabel; Martínez-Fernández, Verónica; Fernández-Pevida, Antonio; Cuevas-Bermúdez, Abel; Martín-Expósito, Manuel; Chávez, Sebastián; de la Cruz, Jesús; Navarro, Francisco

    2014-09-01

    Bud27, the yeast orthologue of human URI/RMP, is a member of the prefoldin-like family of ATP-independent molecular chaperones. It has recently been shown to mediate the assembly of the three RNA polymerases in an Rpb5-dependent manner. In this work, we present evidence of Bud27 modulating RNA pol II transcription elongation. We show that Bud27 associates with RNA pol II phosphorylated forms (CTD-Ser5P and CTD-Ser2P), and that its absence affects RNA pol II occupancy of transcribed genes. We also reveal that Bud27 associates in vivo with the Sth1 component of the chromatin remodeling complex RSC and mediates its association with RNA pol II. Our data suggest that Bud27, in addition of contributing to Rpb5 folding within the RNA polymerases, also participates in the correct assembly of other chromatin-associated protein complexes, such as RSC, thereby modulating their activity.

  19. Transcriptome Profiling of Tiller Buds Provides New Insights into PhyB Regulation of Tillering and Indeterminate Growth in Sorghum.

    Science.gov (United States)

    Kebrom, Tesfamichael H; Mullet, John E

    2016-04-01

    Phytochrome B (phyB) enables plants to modify shoot branching or tillering in response to varying light intensities and ratios of red and far-red light caused by shading and neighbor proximity. Tillering is inhibited in sorghum genotypes that lack phytochrome B (58M, phyB-1) until after floral initiation. The growth of tiller buds in the first leaf axil of wild-type (100M, PHYB) and phyB-1 sorghum genotypes is similar until 6 d after planting when buds of phyB-1 arrest growth, while wild-type buds continue growing and develop into tillers. Transcriptome analysis at this early stage of bud development identified numerous genes that were up to 50-fold differentially expressed in wild-type/phyB-1 buds. Up-regulation of terminal flower1, GA2oxidase, and TPPI could protect axillary meristems in phyB-1 from precocious floral induction and decrease bud sensitivity to sugar signals. After bud growth arrest in phyB-1, expression of dormancy-associated genes such as DRM1, GT1, AF1, and CKX1 increased and ENOD93, ACCoxidase, ARR3/6/9, CGA1, and SHY2 decreased. Continued bud outgrowth in wild-type was correlated with increased expression of genes encoding a SWEET transporter and cell wall invertases. The SWEET transporter may facilitate Suc unloading from the phloem to the apoplast where cell wall invertases generate monosaccharides for uptake and utilization to sustain bud outgrowth. Elevated expression of these genes was correlated with higher levels of cytokinin/sugar signaling in growing buds of wild-type plants. PMID:26893475

  20. A novel minimal in vitro system for analyzing HIV-1 Gag mediated budding

    CERN Document Server

    Gui, Dong; Xu, Jun; Zandi, Roya; Gill, Sarjeet; Huang, I-Chueh; Rao, A L N; Mohideen, Umar

    2013-01-01

    A biomimetic minimalist model membrane is used to study the mechanism and kinetics of the in vitro HIV-1 Gag budding from a giant unilamellar vesicle (GUV). The real time interaction of the Gag, RNA and lipid leading to the formation of minivesicles is measured in real time using confocal microscopy. The Gag is found to lead to resolution limited punctae on the lipid membranes of the GUV. The introduction of the Gag to a GUV solution containing RNA led to the budding of minivesicles on the inside surface of the GUV. The diameter of the GUV decreased due to the bud formation. The corresponding rate of decrease of the GUV diameter was found to be linear in time. The bud formation and the decrease in GUV size were found to be proportional to the Gag concentration. The method is promising and will allow the systematic study of the dynamics of assembly of immature HIV and help classify the hierarchy of factors that impact the Gag protein initiated assembly of retroviruses such as HIV. The GUV system might also be ...

  1. Perception of photoperiod in individual buds of mature trees regulates leaf-out.

    Science.gov (United States)

    Zohner, Constantin M; Renner, Susanne S

    2015-12-01

    Experimental data on the perception of day length and temperature in dormant temperate zone trees are surprisingly scarce. In order to investigate when and where these environmental signals are perceived, we carried out bagging experiments in which buds on branches of Fagus sylvatica, Aesculus hippocastanum and Picea abies trees were exposed to natural light increase or kept at constant 8-h days from December until June. Parallel experiments used twigs cut from the same trees, harvesting treated and control twigs seven times and then exposing them to 8- or 16-h days in a glasshouse. Under 8-h days, budburst in Fagus outdoors was delayed by 41 d and in Aesculus by 4 d; in Picea, day length had no effect. Buds on nearby branches reacted autonomously, and leaf primordia only reacted to light cues in late dormancy after accumulating warm days. Experiments applying different wavelength spectra and high-resolution spectrometry to buds indicate a phytochrome-mediated photoperiod control. By demonstrating local photoperiodic control of buds, revealing the time when these signals are perceived, and showing the interplay between photoperiod and chilling, this study contributes to improved modelling of the impact of climate warming on photosensitive species. PMID:26096967

  2. Autocrine and paracrine roles for ATP and serotonin in mouse taste buds.

    Science.gov (United States)

    Huang, Yijen A; Dando, Robin; Roper, Stephen D

    2009-11-01

    Receptor (type II) taste bud cells secrete ATP during taste stimulation. In turn, ATP activates adjacent presynaptic (type III) cells to release serotonin (5-hydroxytryptamine, or 5-HT) and norepinephrine (NE). The roles of these neurotransmitters in taste buds have not been fully elucidated. Here we tested whether ATP or 5-HT exert feedback onto receptor (type II) cells during taste stimulation. Our previous studies showed NE does not appear to act on adjacent taste bud cells, or at least on receptor cells. Our data show that 5-HT released from presynaptic (type III) cells provides negative paracrine feedback onto receptor cells by activating 5-HT(1A) receptors, inhibiting taste-evoked Ca(2+) mobilization in receptor cells, and reducing ATP secretion. The findings also demonstrate that ATP exerts positive autocrine feedback onto receptor (type II) cells by activating P2Y1 receptors and enhancing ATP secretion. These results begin to sort out how purinergic and aminergic transmitters function within the taste bud to modulate gustatory signaling in these peripheral sensory organs.

  3. Whole lifespan microscopic observation of budding yeast aging through a microfluidic dissection platform

    NARCIS (Netherlands)

    Lee, Sung Sik; Avalos Vizcarra, Ima; Huberts, Daphne H E W; Lee, Luke P; Heinemann, Matthias

    2012-01-01

    Important insights into aging have been generated with the genetically tractable and short-lived budding yeast. However, it is still impossible today to continuously track cells by high-resolution microscopic imaging (e.g., fluorescent imaging) throughout their entire lifespan. Instead, the field st

  4. Continuous High-resolution Microscopic Observation of Replicative Aging in Budding Yeast

    NARCIS (Netherlands)

    Huberts, Daphne H. E. W.; Janssens, Georges E.; Lee, Sung Sik; Vizcarra, Ima Avalos; Heinemann, Matthias

    2013-01-01

    We demonstrate the use of a simple microfluidic setup, in which single budding yeast cells can be tracked throughout their entire lifespan. The microfluidic chip exploits the size difference between mother and daughter cells using an array of micropads. Upon loading, cells are trapped underneath the

  5. Buds of Parenting in Emerging Adult Males: What We Learned from Our Parents

    Science.gov (United States)

    Scharf, Miri; Mayseless, Ofra

    2011-01-01

    The authors examine the precursors of parenting buds (representations regarding parenting before actual parenting) by following 60 men from adolescence to emerging adulthood. Quality of relationships with parents, and attachment representations (state of mind with respect to attachment and attachment styles) assessed in adolescence, contribute to…

  6. Kenny, Bud, and Now Luther! Using Curtis' Books in the Classroom

    Science.gov (United States)

    de Beck, Sharon M.

    2005-01-01

    Christopher Paul Curtis is an internationally known author of books loved by young adult readers. "The Watsons Go to Birmingham--1963" and "Bud, Not Buddy" have now been joined by "Bucking the Sarge," Curtis' newest book. This article will share information about each of the texts, as well as how they can be used in…

  7. Perception of photoperiod in individual buds of mature trees regulates leaf-out.

    Science.gov (United States)

    Zohner, Constantin M; Renner, Susanne S

    2015-12-01

    Experimental data on the perception of day length and temperature in dormant temperate zone trees are surprisingly scarce. In order to investigate when and where these environmental signals are perceived, we carried out bagging experiments in which buds on branches of Fagus sylvatica, Aesculus hippocastanum and Picea abies trees were exposed to natural light increase or kept at constant 8-h days from December until June. Parallel experiments used twigs cut from the same trees, harvesting treated and control twigs seven times and then exposing them to 8- or 16-h days in a glasshouse. Under 8-h days, budburst in Fagus outdoors was delayed by 41 d and in Aesculus by 4 d; in Picea, day length had no effect. Buds on nearby branches reacted autonomously, and leaf primordia only reacted to light cues in late dormancy after accumulating warm days. Experiments applying different wavelength spectra and high-resolution spectrometry to buds indicate a phytochrome-mediated photoperiod control. By demonstrating local photoperiodic control of buds, revealing the time when these signals are perceived, and showing the interplay between photoperiod and chilling, this study contributes to improved modelling of the impact of climate warming on photosensitive species.

  8. Gall mite inspection on dormant black currant buds using machine vision

    DEFF Research Database (Denmark)

    Nielsen, M. R.; Stigaard Laursen, Morten; Jonassen, M. S.;

    2013-01-01

    This paper presents a novel machine vision-based approach detecting and mapping gall mite infection in dormant buds on black currant bushes. A vehicle was fitted with four cameras and RTK-GPS. Results compared automatic detection to human decisions based on the images, and by mapping the results ...

  9. Destinace České Budějovice na trhu cestovního ruchu

    OpenAIRE

    DUŠEK, Pavel

    2013-01-01

    Main target of this bachelor thesis was to evaluate the primary and secondary destination offers of České Budějovice based on situational analysis, identification of competitive destinations and position in the market with tourism. At the same time establishing a competitive advantage of this destination and offering a way of enforcing.

  10. Effect of floral bud reduction on flower longevity in Asiatic hybrids lilies.

    NARCIS (Netherlands)

    Meulen-Muisers, van der J.J.M.; Oeveren, van J.C.; Sandbrink, J.M.; Tuyl, van J.M.

    1995-01-01

    Floral bud abortion was found to be an undesirable source of non-genetic variation in breeding trials directed on the improvement of individual flower longevity in Asiatic hybrid lilies. It increased the longevity of the remaining flowers of the inflorescence. A similar response was found after elim

  11. Reproductive effects of lipid soluble components of Syzygium aromaticum flower bud in male mice

    Directory of Open Access Journals (Sweden)

    Raghav Kumar Mishra

    2013-01-01

    Full Text Available Background: The flower buds of Syzygium aromaticum (clove have been used in indigenous medicines for the treatment of male sexual disorders in Indian subcontinent. Objective: To evaluate the effect of Syzygium aromaticum flower bud on male reproduction, using Parkes (P strain mice as animal model. Materials and Methods: Mice were orally administered lipid soluble components of Syzygium aromaticum flower bud in doses of 15, 30, and 60 mg/kg body weight for 35 days, and several male reproductive endpoints were evaluated. Results: Treatment with lower dose (15 mg of Syzygium increased the motility of sperm and stimulated the secretory activities of epididymis and seminal vesicle, while higher doses (30 and 60 mg had adverse effects on sperm dynamics of cauda epididymidis and on the secretory activities of epididymis and seminal vesicle. Libido was not affected in treated males; however, a significant decrease in litter in females sired by males treated with higher doses of Syzygium was recorded. Conclusion: Treatment with Syzygium aromaticum flower bud causes dose-dependent biphasic effect on male reproductive indices in P mice; lower dose of Syzygium appears stimulatory, while the higher doses have adverse effect on male reproduction. The results suggest that the lower dose of Syzygium may have androgenic effect, but further studies are needed to support this contention.

  12. Mutation breeding in sweet potato with fast neutrons by inducing hypocotyl adventitious buds

    International Nuclear Information System (INIS)

    Hybrid sweet potato seeds were irradiated by fast neutrons. After the irradiation, adventitious buds were induced from the epidermal cells of the hypocotyls of the seedlings. The adventitious buds appeared from the hypocotyls 2 - 3 cm below the cotyledonary node. Detailed observations of cell division and tissue dissection showed that some adventitious buds emerged in 20 - 45 days after seedling emergence. Most of them originated from a single cell. These adventitious bud clones showed a wide mutation spectrum at high frequency, mostly non-chimeric. The mutant characters included the color of leaf, vine, vein, root-tuber skin and the leaf shape, yield, root-tuber dry matter rates, root-tuber numbers, sprouting, early tuber formation, resistance to black rot, root rot, nematodes etc. The results were confirmed in the field; the observed mutant characters were stable and non-chimeric. Such a method of inducing mutations is advantageous for the improvement of economic characters. Actually, there were examples of increasing the dry matter content of the tubers by 1.0 - 5.5%, the yield of tubers increased by 3.6 - 53.8%, and tubers per plant increased by 0.6 - 4.2%. The resistances to black rot, root rot, nematodes etc. were also improved. Three multiple resistance varieties Yanshu 568, Yanshu 571-1, 84-C-2 and two good quality varieties Yanshu 759 and 781 have been selected. (author). 9 refs, 3 tabs

  13. Efficient transmission of Cassava brown streak disease viral pathogens by chip bud grafting

    Science.gov (United States)

    2013-01-01

    Background Techniques to study plant viral diseases under controlled growth conditions are required to fully understand their biology and investigate host resistance. Cassava brown streak disease (CBSD) presents a major threat to cassava production in East Africa. No infectious clones of the causal viruses, Cassava brown streak virus (CBSV) or Ugandan cassava brown streak virus (UCBSV) are available, and mechanical transmission to cassava is not effective. An improved method for transmission of the viruses, both singly and as co-infections has been developed using bud grafts. Findings Axillary buds from CBSD symptomatic plants infected with virulent isolates of CBSV and UCBSV were excised and grafted onto 6–8 week old greenhouse-grown, disease-free cassava plants of cultivars Ebwanateraka, TME204 and 60444. Plants were assessed visually for development of CBSD symptoms and by RT-PCR for presence of the viruses in leaf and storage root tissues. Across replicated experiments, 70-100% of plants inoculated with CBSV developed CBSD leaf and stem symptoms 2–6 weeks after bud grafting. Infected plants showed typical, severe necrotic lesions in storage roots at harvest 12–14 weeks after graft inoculation. Sequential grafting of buds from plants infected with UCBSV followed 10–14 days later by buds carrying CBSV, onto the same test plant, resulted in 100% of the rootstocks becoming co-infected with both pathogens. This dual transmission rate was greater than that achieved by simultaneous grafting with UCBSV and CBSV (67%), or when grafting first with CBSV followed by UCBSV (17%). Conclusions The bud grafting method described presents an improved tool for screening cassava germplasm for resistance to CBSD causal viruses, and for studying pathogenicity of this important disease. Bud grafting provides new opportunities compared to previously reported top and side grafting systems. Test plants can be inoculated as young, uniform plants of a size easily handled in a

  14. Adenosine enhances sweet taste through A2B receptors in the taste bud.

    Science.gov (United States)

    Dando, Robin; Dvoryanchikov, Gennady; Pereira, Elizabeth; Chaudhari, Nirupa; Roper, Stephen D

    2012-01-01

    Mammalian taste buds use ATP as a neurotransmitter. Taste Receptor (type II) cells secrete ATP via gap junction hemichannels into the narrow extracellular spaces within a taste bud. This ATP excites primary sensory afferent fibers and also stimulates neighboring taste bud cells. Here we show that extracellular ATP is enzymatically degraded to adenosine within mouse vallate taste buds and that this nucleoside acts as an autocrine neuromodulator to selectively enhance sweet taste. In Receptor cells in a lingual slice preparation, Ca(2+) mobilization evoked by focally applied artificial sweeteners was significantly enhanced by adenosine (50 μM). Adenosine had no effect on bitter or umami taste responses, and the nucleoside did not affect Presynaptic (type III) taste cells. We also used biosensor cells to measure transmitter release from isolated taste buds. Adenosine (5 μM) enhanced ATP release evoked by sweet but not bitter taste stimuli. Using single-cell reverse transcriptase (RT)-PCR on isolated vallate taste cells, we show that many Receptor cells express the adenosine receptor, Adora2b, while Presynaptic (type III) and Glial-like (type I) cells seldom do. Furthermore, Adora2b receptors are significantly associated with expression of the sweet taste receptor subunit, Tas1r2. Adenosine is generated during taste stimulation mainly by the action of the ecto-5'-nucleotidase, NT5E, and to a lesser extent, prostatic acid phosphatase. Both these ecto-nucleotidases are expressed by Presynaptic cells, as shown by single-cell RT-PCR, enzyme histochemistry, and immunofluorescence. Our findings suggest that ATP released during taste reception is degraded to adenosine to exert positive modulation particularly on sweet taste.

  15. Mapping of Candidate Genes Involved in Bud Dormancy and Flowering Time in Sweet Cherry (Prunus avium.

    Directory of Open Access Journals (Sweden)

    Sophie Castède

    Full Text Available The timing of flowering in perennial plants is crucial for their survival in temperate climates and is regulated by the duration of bud dormancy. Bud dormancy release and bud break depend on the perception of cumulative chilling during endodormancy and heat during the bud development. The objectives of this work were to identify candidate genes involved in dormancy and flowering processes in sweet cherry, their mapping in two mapping progenies 'Regina' × 'Garnet' and 'Regina' × 'Lapins', and to select those candidate genes which co-localized with quantitative trait loci (QTLs associated with temperature requirements for bud dormancy release and flowering. Based on available data on flowering processes in various species, a list of 79 candidate genes was established. The peach and sweet cherry orthologs were identified and primers were designed to amplify sweet cherry candidate gene fragments. Based on the amplified sequences of the three parents of the mapping progenies, SNPs segregations in the progenies were identified. Thirty five candidate genes were genetically mapped in at least one of the two progenies and all were in silico mapped. Co-localization between candidate genes and QTLs associated with temperature requirements and flowering date were identified for the first time in sweet cherry. The allelic composition of the candidate genes located in the major QTL for heat requirements and flowering date located on linkage group 4 have a significant effect on these two traits indicating their potential use for breeding programs in sweet cherry to select new varieties adapted to putative future climatic conditions.

  16. THE BUD BREAK PROCESS AND ITS VARIATION AMONG LOCAL POPULATIONS OF BOREAL BLACK SPRUCE

    Directory of Open Access Journals (Sweden)

    Sergio eRossi

    2014-10-01

    Full Text Available Phenology of local populations can exhibit adaptations to the current environmental conditions resulting from a close interaction between climate and genotype. The bud break process and its variations among populations were analysed in greenhouse by monitoring the growth resumption in black spruce [Picea mariana (Mill. BSP] seedlings originating from seeds of five stands across the closed boreal forest in Quebec, Canada. Bud break lasted 15 days and occurred earlier and quicker in northern provenances. Provenance explained between 10.2 and 32.3% of the variance in bud break, while the families accounted for a smaller but still significant part of the variance. The late occurrence of one phenological phase corresponded to a delayed occurrence of the others according to linear relationships. A causal model was proposed in the form of a chain of events with each phase of bud break being related to the previous and successive one, while no link was observed between non-adjacent phases. The adaptation of black spruce populations along the latitudinal gradient points towards a strategy based on rapid physiological processes triggered by temperature increase inducing high metabolic activity. The variation observed in bud break reflects an evolutionary trade-off between maximization of security and taking advantage of the short growing season. This work provides evidence of the phenological adaptations of black spruce to its local environmental conditions while retaining sizeable genetic diversity within populations. Because of the multigenic nature of phenology, this diversity should provide some raw material for adaptation to changing local environmental conditions.

  17. The bud break process and its variation among local populations of boreal black spruce

    Science.gov (United States)

    Rossi, Sergio; Bousquet, Jean

    2014-01-01

    Phenology of local populations can exhibit adaptations to the current environmental conditions resulting from a close interaction between climate and genotype. The bud break process and its variations among populations were analyzed in greenhouse by monitoring the growth resumption in black spruce [Picea mariana (Mill.) BSP] seedlings originating from seeds of five stands across the closed boreal forest in Quebec, Canada. Bud break lasted 15 days and occurred earlier and quicker in northern provenances. Provenance explained between 10.2 and 32.3% of the variance in bud break, while the families accounted for a smaller but still significant part of the variance. The late occurrence of one phenological phase corresponded to a delayed occurrence of the others according to linear relationships. A causal model was proposed in the form of a chain of events with each phase of bud break being related to the previous and successive one, while no link was observed between non-adjacent phases. The adaptation of black spruce populations along the latitudinal gradient points toward a strategy based on rapid physiological processes triggered by temperature increase inducing high metabolic activity. The variation observed in bud break reflects an evolutionary trade-off between maximization of security and taking advantage of the short growing season. This work provides evidence of the phenological adaptations of black spruce to its local environmental conditions while retaining sizeable genetic diversity within populations. Because of the multigenic nature of phenology, this diversity should provide some raw material for adaptation to changing local environmental conditions. PMID:25389430

  18. Mapping of Candidate Genes Involved in Bud Dormancy and Flowering Time in Sweet Cherry (Prunus avium).

    Science.gov (United States)

    Castède, Sophie; Campoy, José Antonio; Le Dantec, Loïck; Quero-García, José; Barreneche, Teresa; Wenden, Bénédicte; Dirlewanger, Elisabeth

    2015-01-01

    The timing of flowering in perennial plants is crucial for their survival in temperate climates and is regulated by the duration of bud dormancy. Bud dormancy release and bud break depend on the perception of cumulative chilling during endodormancy and heat during the bud development. The objectives of this work were to identify candidate genes involved in dormancy and flowering processes in sweet cherry, their mapping in two mapping progenies 'Regina' × 'Garnet' and 'Regina' × 'Lapins', and to select those candidate genes which co-localized with quantitative trait loci (QTLs) associated with temperature requirements for bud dormancy release and flowering. Based on available data on flowering processes in various species, a list of 79 candidate genes was established. The peach and sweet cherry orthologs were identified and primers were designed to amplify sweet cherry candidate gene fragments. Based on the amplified sequences of the three parents of the mapping progenies, SNPs segregations in the progenies were identified. Thirty five candidate genes were genetically mapped in at least one of the two progenies and all were in silico mapped. Co-localization between candidate genes and QTLs associated with temperature requirements and flowering date were identified for the first time in sweet cherry. The allelic composition of the candidate genes located in the major QTL for heat requirements and flowering date located on linkage group 4 have a significant effect on these two traits indicating their potential use for breeding programs in sweet cherry to select new varieties adapted to putative future climatic conditions. PMID:26587668

  19. The role of spatially controlled cell proliferation in limb bud morphogenesis.

    Directory of Open Access Journals (Sweden)

    Bernd Boehm

    Full Text Available Although the vertebrate limb bud has been studied for decades as a model system for spatial pattern formation and cell specification, the cellular basis of its distally oriented elongation has been a relatively neglected topic by comparison. The conventional view is that a gradient of isotropic proliferation exists along the limb, with high proliferation rates at the distal tip and lower rates towards the body, and that this gradient is the driving force behind outgrowth. Here we test this hypothesis by combining quantitative empirical data sets with computer modelling to assess the potential role of spatially controlled proliferation rates in the process of directional limb bud outgrowth. In particular, we generate two new empirical data sets for the mouse hind limb--a numerical description of shape change and a quantitative 3D map of cell cycle times--and combine these with a new 3D finite element model of tissue growth. By developing a parameter optimization approach (which explores spatial patterns of tissue growth our computer simulations reveal that the observed distribution of proliferation rates plays no significant role in controlling the distally extending limb shape, and suggests that directional cell activities are likely to be the driving force behind limb bud outgrowth. This theoretical prediction prompted us to search for evidence of directional cell orientations in the limb bud mesenchyme, and we thus discovered a striking highly branched and extended cell shape composed of dynamically extending and retracting filopodia, a distally oriented bias in Golgi position, and also a bias in the orientation of cell division. We therefore provide both theoretical and empirical evidence that limb bud elongation is achieved by directional cell activities, rather than a PD gradient of proliferation rates.

  20. The bud break process and its variation among local populations of boreal black spruce.

    Science.gov (United States)

    Rossi, Sergio; Bousquet, Jean

    2014-01-01

    Phenology of local populations can exhibit adaptations to the current environmental conditions resulting from a close interaction between climate and genotype. The bud break process and its variations among populations were analyzed in greenhouse by monitoring the growth resumption in black spruce [Picea mariana (Mill.) BSP] seedlings originating from seeds of five stands across the closed boreal forest in Quebec, Canada. Bud break lasted 15 days and occurred earlier and quicker in northern provenances. Provenance explained between 10.2 and 32.3% of the variance in bud break, while the families accounted for a smaller but still significant part of the variance. The late occurrence of one phenological phase corresponded to a delayed occurrence of the others according to linear relationships. A causal model was proposed in the form of a chain of events with each phase of bud break being related to the previous and successive one, while no link was observed between non-adjacent phases. The adaptation of black spruce populations along the latitudinal gradient points toward a strategy based on rapid physiological processes triggered by temperature increase inducing high metabolic activity. The variation observed in bud break reflects an evolutionary trade-off between maximization of security and taking advantage of the short growing season. This work provides evidence of the phenological adaptations of black spruce to its local environmental conditions while retaining sizeable genetic diversity within populations. Because of the multigenic nature of phenology, this diversity should provide some raw material for adaptation to changing local environmental conditions. PMID:25389430

  1. Cell-to-cell communication in intact taste buds through ATP signalling from pannexin 1 gap junction hemichannels.

    Science.gov (United States)

    Dando, Robin; Roper, Stephen D

    2009-12-15

    Isolated taste cells, taste buds and strips of lingual tissue from taste papillae secrete ATP upon taste stimulation. Taste bud receptor (Type II) cells have been identified as the source of ATP secretion. Based on studies on isolated taste buds and single taste cells, we have postulated that ATP secreted from receptor cells via pannexin 1 hemichannels acts within the taste bud to excite neighbouring presynaptic (Type III) cells. This hypothesis, however, remains to be tested in intact tissues. In this report we used confocal Ca(2+) imaging and lingual slices containing intact taste buds to test the hypothesis of purinergic signalling between taste cells in a more integral preparation. Incubating lingual slices with apyrase reversibly blocked cell-to-cell communication between receptor cells and presynaptic cells, consistent with ATP being the transmitter. Inhibiting pannexin 1 gap junction hemichannels with CO(2)-saturated buffer or probenecid significantly reduced cell-cell signalling between receptor cells and presynaptic cells. In contrast, anandamide, a blocker of connexin gap junction channels, had no effect of cell-to-cell communication in taste buds. These findings are consistent with the model for peripheral signal processing via ATP and pannexin 1 hemichannels in mammalian taste buds.

  2. Epigenetic drug 5-azacytidine impairs proliferation of rat limb buds in an organotypic model-system in vitro

    Science.gov (United States)

    Mužić, Vedrana; Katušić Bojanac, Ana; Jurić-Lekić, Gordana; Himelreich, Marta; Tupek, Katarina; Šerman, Ljiljana; Marn, Nina; Sinčić, Nino; Vlahović, Maja; Bulić-Jakuš, Floriana

    2013-01-01

    Aim To establish an organotypic in vitro model of limb bud development to verify whether epigenetic drug and teratogen 5-azacytidine (5azaC) has an effect on limb buds independent of its effects on the placenta. Methods Fischer strain rat fore- and hindlimb buds were microsurgically isolated from 13 days old embryos and cultivated in vitro for two weeks at the air-liquid interface in Eagle's minimum essential medium (MEM) with 50% rat serum. 30 µmol of 5azaC was added to the fresh medium. Overall growth was measured by an ocular micrometer. Routine histology, immunohistochemical detection of the proliferating cell nuclear antigen (PCNA), and stereological quantification of PCNA expression were performed. Results At four time points, significantly lower overall growth was detected for fore- and hindlimb bud explants cultivated with 5azaC in comparison to controls. After the culture period, numerical density of the PCNA signal for both types of limb buds was lower than for controls (P < 0.001). Limb buds were initially covered by immature epithelium and contained mesenchyme, myotubes, single hemangioblasts, hemangioblast aggregates, blood islands, and capillaries. Regardless of the treatment, cartilage and epidermis differentiated, but cells and structures typical for vasculogenesis disappeared. Conclusion Our findings, obtained outside of the maternal organism, stress the importance of compromised cell proliferation for 5azaC impact on limb buds. This investigation points to the necessity to establish alternatives to in vivo research on animals using teratogenic agents. PMID:24170728

  3. 荞麦芽的抗氧化活性研究%Study on Antioxidant Activity of Buckwheat Buds

    Institute of Scientific and Technical Information of China (English)

    孙国娟; 桂英; 刘笑笑; 崔泰花; 崔承弼

    2012-01-01

    In order to research the antioxidant activity of buckwheat buds, the experiment with different growth stages of buckwheat buds as the object of study by 70% ethanol solvent extracting total flavonoids of the different growth stages of sweet buckwheat buds and bitter buckwheat buds were studied. The effect on eliminating DPPH from the extract of buckwheat buds were determined by colorimetry. The results indicated that the different varieties of buckwheat buds had certain antioxidant capacity. And the antioxidant capacity of bitter buckwheat was significantly higher than sweet buckwheat. The ability of buckwheat buds on DPPH free radical clearance was the highest on the 14th d of growing period. Bitter buckwheat buds achieved 80% above, sweet buckwheat buds in more than 50%, after a few days it gadually declined within a narrow range.%为了研究荞麦芽的抗氧化活性,以不同生长阶段的荞麦芽为研究对象,采用70%乙醇为溶剂提取了不同生长阶段甜荞麦芽和苦荞麦芽的总黄酮,利用比色法测定了荞麦芽提取物对DPPH自由基的清除率。结果表明,不同品种的荞麦芽苗均有一定的抗氧化能力,且苦荞的抗氧化能力明显高于甜荞。并在第14d时荞麦芽提取物对DPPH自由基的清除能力最高,苦荞麦芽达到80%以上,甜荞麦芽在50%以上,以后几天略有下降。

  4. Epigenetic drug 5-azacytidine impairs proliferation of rat limb buds in an organotypic model-system in vitro

    OpenAIRE

    Mužić, Vedrana; Katušić Bojanac, Ana; Jurić-Lekić, Gordana; Himelreich, Marta; Tupek, Katarina; Šerman, Ljiljana; Marn, Nina; Sinčić, Nino; Vlahović, Maja; Bulić-Jakuš, Floriana

    2013-01-01

    Aim To establish an organotypic in vitro model of limb bud development to verify whether epigenetic drug and teratogen 5-azacytidine (5azaC) has an effect on limb buds independent of its effects on the placenta. Methods Fischer strain rat fore- and hindlimb buds were microsurgically isolated from 13 days old embryos and cultivated in vitro for two weeks at the air-liquid interface in Eagle’s minimum essential medium (MEM) with 50% rat serum. 30 μmol of 5azaC was added ...

  5. Effects of 6-BA and MET on the induction of adventitious buds from rice seedlings and its application

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@Different levels of 6-BA (6-Benezyla-mino purine) and MET (Paclobutra-zol) were added into MS media to culture the mature seeds of six varieties and two F2 populations of Nanjing11 ( indica ) / Balilla (japoni-ca) and Dular( indica)/Balilla( japonica) . Results were as follows:1) 6-BA was effective in inducing the formation of adventitious buds (Table 1).Only using MET could not induce adventitious buds formation. Compared with the use of 6-BA only, it was more useful for inducing the adventitious buds by using MET and 6-BA together, especially for japonica rice.

  6. Annexin VI-mediated Loss of Spectrin during Coated Pit Budding Is Coupled to Delivery of LDL to Lysosomes

    OpenAIRE

    Kamal, Adeela; Ying, Yun-shu; Anderson, Richard G. W.

    1998-01-01

    Previously we reported that annexin VI is required for the budding of clathrin-coated pits from human fibroblast plasma membranes in vitro. Here we show that annexin VI bound to the NH2-terminal 28-kD portion of membrane spectrin is as effective as cytosolic annexin VI in supporting coated pit budding. Annexin VI–dependent budding is accompanied by the loss of ∼50% of the spectrin from the membrane and is blocked by the cysteine protease inhibitor N-acetyl-leucyl-leucyl-norleucinal (ALLN). In...

  7. Assessment of factors affecting in vitro shoot regeneration from axillary bud explant of Camptotheca acuminata

    Institute of Scientific and Technical Information of China (English)

    WANG Hui-Mei; ZU Yuan-Gang; DONG Feng-Li; ZHAO Xiao-Ju

    2005-01-01

    Axillary buds from 3-yr.-old seedlings of Camptotheca acuminata in the greenhouse were cultured on the different basal media with different concentrations of growth regulators for shoot regeneration for studying the effects of different basal media, different concentrations of growth regulators (BA or TDZ), sucrose, agar and pH value on shoot regeneration from axillary bud. The results showed that B5 and WPM media were the optimal basal media and the optimal phyotohormone was BA of 1.0 mg/L or TDZ of 0.1mg/L; The concentrations of sucrose of 30g/L and agar of 6g/L were most suitable for the shoot regeneration; pH value from 5.8 to 6.6 were broadly effective, but the best at pH 5.8.

  8. ALIX is recruited temporarily into HIV-1 budding sites at the end of gag assembly.

    Directory of Open Access Journals (Sweden)

    Pei-I Ku

    Full Text Available Polymerization of Gag on the inner leaflet of the plasma membrane drives the assembly of Human Immunodeficiency Virus 1 (HIV-1. Gag recruits components of the endosomal sorting complexes required for transport (ESCRT to facilitate membrane fission and virion release. ESCRT assembly is initiated by recruitment of ALIX and TSG101/ESCRT-I, which bind directly to the viral Gag protein and then recruit the downstream ESCRT-III and VPS4 factors to complete the budding process. In contrast to previous models, we show that ALIX is recruited transiently at the end of Gag assembly, and that most ALIX molecules are recycled into the cytosol as the virus buds, although a subset remains within the virion. Our experiments imply that ALIX is recruited to the neck of the assembling virion and is mostly recycled after virion release.

  9. What is the role of metabolic hormones in taste buds of the tongue.

    Science.gov (United States)

    Cai, Huan; Maudsley, Stuart; Martin, Bronwen

    2014-01-01

    Gustation is one of the important chemical senses that guides the organism to identify nutrition while avoiding toxic chemicals. An increasing number of metabolic hormones and/or hormone receptors have been identified in the taste buds of the tongue and are involved in modulating taste perception. The gustatory system constitutes an additional endocrine regulatory locus that affects food intake, and in turn whole-body energy homeostasis. Here we provide an overview of the main metabolic hormones known to be present in the taste buds of the tongue; discuss their potential functional roles in taste perception and energy homeostasis and how their functional integrity is altered in the metabolic imbalance status (obesity and diabetes) and aging process. Better understanding of the functional roles of metabolic hormones in flavor perception as well as the link between taste perception and peripheral metabolism may be vital for developing strategies to promote healthier eating and prevent obesity or lifestyle-related disorders.

  10. Fimbrin phosphorylation by metaphase Cdk1 regulates actin cable dynamics in budding yeast.

    Science.gov (United States)

    Miao, Yansong; Han, Xuemei; Zheng, Liangzhen; Xie, Ying; Mu, Yuguang; Yates, John R; Drubin, David G

    2016-01-01

    Actin cables, composed of actin filament bundles nucleated by formins, mediate intracellular transport for cell polarity establishment and maintenance. We previously observed that metaphase cells preferentially promote actin cable assembly through cyclin-dependent kinase 1 (Cdk1) activity. However, the relevant metaphase Cdk1 targets were not known. Here we show that the highly conserved actin filament crosslinking protein fimbrin is a critical Cdk1 target for actin cable assembly regulation in budding yeast. Fimbrin is specifically phosphorylated on threonine 103 by the metaphase cyclin-Cdk1 complex, in vivo and in vitro. On the basis of conformational simulations, we suggest that this phosphorylation stabilizes fimbrin's N-terminal domain, and modulates actin filament binding to regulate actin cable assembly and stability in cells. Overall, this work identifies fimbrin as a key target for cell cycle regulation of actin cable assembly in budding yeast, and suggests an underlying mechanism.

  11. [Antiradical properties of essential oils and extracts from clove bud and pimento].

    Science.gov (United States)

    Misharina, T A; Alinkina, E S; Medvedeva, I B

    2015-01-01

    The antiradical properties of essential oils and extracts from the clove bud (Eugenia caryophyllata Thumb.) and berries of tree (Pimenta dioica (L.) Meriff) were studied and compared with the properties of synthetic antioxidant ionol (2,6-ditret-butyl-4-hydroxytoluene, BHT) in model reactions with the stable free 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. The essential oils of clove bud and pimento had qualitatively close composition of the main components but differed by their quantitative content. In the studied samples, eugenol was the main compound with high antiradical activity. The reaction rates of essential oils and extracts with the DPPH radical were practically the same for essential oils and twice the reaction rate of BHT. The values of antiradical efficiency (AE) were also close for essential oils and were twice that for extracts and ionol. A synergetic action of components in the essential oil and extract of pimento on antiradical efficiency values was found. PMID:25842910

  12. Gravity-dependent regulation of red light induced moss protonemata branching and gametophore bud formation

    Science.gov (United States)

    Ripetskyj, R. T.; Kit, N. A.

    Isolated leafy shoots of the moss Pottia intermedia positioned horizontally on the agar surface in vertically oriented petri dishes regenerate unbranching negatively gravitropic protonemata on upper side of the regenerant. Gravity determines the site of regeneration not the process itself. White light of low intensity unsufficient to induce positive phototropism of dark-grown protonemata can, however, provoke their branching and gametophore bud formation (Ripetskyj et al., 1998; 1999). The presented experiments have been carried out with red light in Biological Research in Canisters/Light Emitting Diode (BRIC/LED) hardware developed at Kennedy Space Center, USA. Seven-day-old dark-grown negatively gravitropic secondary P. intermedia protonemata were positioned differently with respect to gravity vector and to the source of red light of low, 1 or 2 μ mol\\cdot m-2\\cdot s-1, intensities. The light induced intensive branching of the protonemata and gametophore bud formation initiation site of both processes as well as the direction of growth of branches and buds being depent on the position of protonemata with respect to gravity and light vectors. Vertically positioned, i.e. ungravistimulated, dark grown protonemata illuminated from one side with red light of 2 μ mol\\cdot m-2\\cdot s-1 intensity produced 96,9 ± 2,2% of side branches and buds growing directly towards the light source from the lit protonema side. Horizontally disposed protonemata irradiated from below with red light of the same intensity regenerate 31,7 ± 3,9% of branches and buds on the upper, i.e. shaded protonemata side, the upward growth of which should undoubtedly be determined by gravity. In vertically disposed protonemata illuminated with red light of 1 μ mol\\cdot m-2\\cdot s-1 intensity from aside 31,9 ± 5,5% of side branches and buds arised on shaded protonema side and grew away from the light. Illumination of the protonemata in horizontal position from below increased the number of

  13. Whole lifespan microscopic observation of budding yeast aging through a microfluidic dissection platform

    OpenAIRE

    Lee, Sung Sik; Avalos Vizcarra, Ima; Huberts, Daphne H. E. W.; Lee, Luke P; Heinemann, Matthias

    2012-01-01

    Important insights into aging have been generated with the genetically tractable and short-lived budding yeast. However, it is still impossible today to continuously track cells by high-resolution microscopic imaging (e.g., fluorescent imaging) throughout their entire lifespan. Instead, the field still needs to rely on a 50-y-old laborious and time-consuming method to assess the lifespan of yeast cells and to isolate differentially aged cells for microscopic snapshots via manual dissection of...

  14. CONVENTIONAL PROPAGATION OF SEVERAL AGLAONEMA ACCESSIONS USING SPLIT SINGLE-BUD STEM CUTTING

    OpenAIRE

    Budiarto, Kurniawan

    2011-01-01

    Aglaonema was usually propagated by seeds, sucker separations and stem cuttings. For most cultivars, stem cuttings were still considered the most efficient method. The practice might increase propagation efficiency in Aglaonema production. The research was conducted to find out the effects of nodal age on the rooting capacity and cutting performance of several Aglaonema accessions using split single-bud stem cuttings. The experiment was carried out at the Indonesian Ornamental ...

  15. Effect of gamma rays on vegetative buds of physalis peruviana l

    OpenAIRE

    Caro Melgarejo, Diana Patricia; Estupiñán-Rincón, Sandra Yaneth; Rache Cardenal, Leidy Yanira; Pacheco-Maldonado, José Constantino

    2013-01-01

    In Colombia, studies carried out to establish modern methods for propagation and selection of plant of ribes gooseberry (Physalis peruviana) are scarce so, it is necessary to explore alternative techniques such as the application of ionizing radiation, which enables greater genetic variability and to obtain seedlings with phenotypic characteristics that could be beneficial to contribute to improve productivity. To induce chromosomal alterations, axillary buds of P. peruviana with 50, 100, :00...

  16. CENP-A exceeds microtubule attachment sites in centromere clusters of both budding and fission yeast

    OpenAIRE

    Coffman, Valerie C.; Wu, Pengcheng; Parthun, Mark R.; Wu, Jian-Qiu

    2011-01-01

    The stoichiometries of kinetochores and their constituent proteins in yeast and vertebrate cells were determined using the histone H3 variant CENP-A, known as Cse4 in budding yeast, as a counting standard. One Cse4-containing nucleosome exists in the centromere (CEN) of each chromosome, so it has been assumed that each anaphase CEN/kinetochore cluster contains 32 Cse4 molecules. We report that anaphase CEN clusters instead contained approximately fourfold more Cse4 in Saccharomyces cerevisiae...

  17. Cell wall and lipid composition of Isosphaera pallida, a budding eubacterium from hot springs.

    OpenAIRE

    Giovannoni, S J; Godchaux, W; Schabtach, E; Castenholz, R W

    1987-01-01

    Isosphaera pallida is an unusual gliding, budding eubacterium recently isolated from North American hot springs. Electron micrographs of ultrathin sections revealed a cell wall atypical of eubacteria: two electrondense layers separated by an electron-transparent layer, with no evident peptidoglycan layer. Growth was not inhibited by penicillin. Cell walls were isolated from sheared cells by velocity sedimentation. The rigid-layer fraction, prepared from cell walls by treatment with boiling 10...

  18. Calcium Regulation of Hemorrhagic Fever Virus Budding: Mechanistic Implications for Host-Oriented Therapeutic Intervention.

    Directory of Open Access Journals (Sweden)

    Ziying Han

    2015-10-01

    Full Text Available Hemorrhagic fever viruses, including the filoviruses (Ebola and Marburg and arenaviruses (Lassa and Junín viruses, are serious human pathogens for which there are currently no FDA approved therapeutics or vaccines. Importantly, transmission of these viruses, and specifically late steps of budding, critically depend upon host cell machinery. Consequently, strategies which target these mechanisms represent potential targets for broad spectrum host oriented therapeutics. An important cellular signal implicated previously in EBOV budding is calcium. Indeed, host cell calcium signals are increasingly being recognized to play a role in steps of entry, replication, and transmission for a range of viruses, but if and how filoviruses and arenaviruses mobilize calcium and the precise stage of virus transmission regulated by calcium have not been defined. Here we demonstrate that expression of matrix proteins from both filoviruses and arenaviruses triggers an increase in host cytoplasmic Ca2+ concentration by a mechanism that requires host Orai1 channels. Furthermore, we demonstrate that Orai1 regulates both VLP and infectious filovirus and arenavirus production and spread. Notably, suppression of the protein that triggers Orai activation (Stromal Interaction Molecule 1, STIM1 and genetic inactivation or pharmacological blockade of Orai1 channels inhibits VLP and infectious virus egress. These findings are highly significant as they expand our understanding of host mechanisms that may broadly control enveloped RNA virus budding, and they establish Orai and STIM1 as novel targets for broad-spectrum host-oriented therapeutics to combat these emerging BSL-4 pathogens and potentially other enveloped RNA viruses that bud via similar mechanisms.

  19. Analisis Histologi Embriogenesis Somatik Dari Apikal Bud Kelapa Sawit (Elaeis guineensis Jacq.) var Tenera

    OpenAIRE

    Meilvana, Tengku Nilayanda

    2014-01-01

    A study of Histological Analysis of Somatic Embryogenesis from Apical Bud of Oil Palm (Elaeis guineensis Jacq.) var. Tenera has been conducted at Plant Tissue Culture Laboratory of FMIPA USU. The experimental design was completely randomized with three levels of 2,4-D concentrations: 110; 120; 130 mg/L and three position of explants: apical, median, and basal. The statistical analysis showed that 120 mg/L of 2,4-D significantly affected the primary and embryogenic calluses initiation. Explant...

  20. Asc1 supports cell-wall integrity near bud sites by a Pkc1 independent mechanism.

    Directory of Open Access Journals (Sweden)

    Daniel Melamed

    Full Text Available BACKGROUND: The yeast ribosomal protein Asc1 is a WD-protein family member. Its mammalian ortholog, RACK1 was initially discovered as a receptor for activated protein C kinase (PKC that functions to maintain the active conformation of PKC and to support its movement to target sites. In the budding yeast though, a connection between Asc1p and the PKC signaling pathway has never been reported. METHODOLOGY/PRINCIPAL FINDINGS: In the present study we found that asc1-deletion mutant (asc1Delta presents some of the hallmarks of PKC signaling mutants. These include an increased sensitivity to staurosporine, a specific Pkc1p inhibitor, and susceptibility to cell-wall perturbing treatments such as hypotonic- and heat shock conditions and zymolase treatment. Microscopic analysis of asc1Delta cells revealed cell-wall invaginations near bud sites after exposure to hypotonic conditions, and the dynamic of cells' survival after this stress further supports the involvement of Asc1p in maintaining the cell-wall integrity during the mid-to late stages of bud formation. Genetic interactions between asc1 and pkc1 reveal synergistic sensitivities of a double-knock out mutant (asc1Delta/pkc1Delta to cell-wall stress conditions, and high basal level of PKC signaling in asc1Delta. Furthermore, Asc1p has no effect on the cellular distribution or redistribution of Pkc1p at optimal or at cell-wall stress conditions. CONCLUSIONS/SIGNIFICANCE: Taken together, our data support the idea that unlike its mammalian orthologs, Asc1p acts remotely from Pkc1p, to regulate the integrity of the cell-wall. We speculate that its role is exerted through translation regulation of bud-site related mRNAs during cells' growth.

  1. Investigations on The Changes of Phenolic Substances During Flower Bud Development in Strawberries

    OpenAIRE

    ÖZEKER, Elmas; TANRISEVER, Ali

    1998-01-01

    The extracts prepared from the samples which were collected weekly intervals betweeen August and December in 1990 and 1992 were analyzed by thin layer chromatography and the changes in phenolic substances during flower bud developmental stages in strawberries were examined. Some differences were determined in terms of phenolic compositions among the developmental stages in the one dimensional chromatograms of young and mature leaves of Pocahontas, Tioga and Yalova-110 strawberry cultivars and...

  2. Integrace Ukrajinců v Českých Budějovicích

    OpenAIRE

    ŠILLEROVÁ, Veronika

    2015-01-01

    The diploma thesis is named Integration of Ukrainians in České Budějovice. The first chapter explains the basic concepts related to the topic of integration of foreigners, such as migration, immigrant or refugee. The second section refers to the concept of integration, which is defined in detail. The third chapter describes the integration policy in the Czech Republic. The fourth chapter is dedicated to legislation and two basic laws governing this issue. The fifth chapter concerns foreigners...

  3. Modulation of taste sensitivity by GLP-1 signaling in taste buds

    OpenAIRE

    Martin, Bronwen; Dotson, Cedrick D; Shin, Yu-Kyong; Ji, Sunggoan; Drucker, Daniel J.; Maudsley, Stuart; Munger, Steven D.

    2009-01-01

    Modulation of sensory function can help animals adjust to a changing external and internal environment. Even so, mechanisms for modulating taste sensitivity are poorly understood. Using immunohistochemical, biochemical and behavioral approaches, we found that the peptide hormone glucagon-like peptide-1 (GLP-1) and its receptor (GLP-1R) are expressed in mammalian taste buds. Furthermore, we found that GLP-1 signaling plays an important role in the modulation of taste sensitivity: GLP-1R knocko...

  4. Generation of micronuclei during interphase by coupling between cytoplasmic membrane blebbing and nuclear budding.

    Directory of Open Access Journals (Sweden)

    Koh-ichi Utani

    Full Text Available Micronucleation, mediated by interphase nuclear budding, has been repeatedly suggested, but the process is still enigmatic. In the present study, we confirmed the previous observation that there are lamin B1-negative micronuclei in addition to the positive ones. A large cytoplasmic bleb was found to frequently entrap lamin B1-negative micronuclei, which were connected to the nucleus by a thin chromatin stalk. At the bottom of the stalk, the nuclear lamin B1 structure appeared broken. Chromatin extrusion through lamina breaks has been referred to as herniation or a blister of the nucleus, and has been observed after the expression of viral proteins. A cell line in which extrachromosomal double minutes and lamin B1 protein were simultaneously visualized in different colors in live cells was established. By using these cells, time-lapse microscopy revealed that cytoplasmic membrane blebbing occurred simultaneously with the extrusion of nuclear content, which generated lamin B1-negative micronuclei during interphase. Furthermore, activation of cytoplasmic membrane blebbing by the addition of fresh serum or camptothecin induced nuclear budding within 1 to 10 minutes, which suggested that blebbing might be the cause of the budding. After the induction of blebbing, the frequency of lamin-negative micronuclei increased. The budding was most frequent during S phase and more efficiently entrapped small extrachromosomal chromatin than the large chromosome arm. Based on these results, we suggest a novel mechanism in which cytoplasmic membrane dynamics pulls the chromatin out of the nucleus through the lamina break. Evidence for such a mechanism was obtained in certain cancer cell lines including human COLO 320 and HeLa. The mechanism could significantly perturb the genome and influence cancer cell phenotypes.

  5. Screening the Budding Yeast Genome Reveals Unique Factors Affecting K2 Toxin Susceptibility

    OpenAIRE

    Elena Servienė; Juliana Lukša; Irma Orentaitė; Lafontaine, Denis L. J.; Jaunius Urbonavičius

    2012-01-01

    BACKGROUND: Understanding how biotoxins kill cells is of prime importance in biomedicine and the food industry. The budding yeast (S. cerevisiae) killers serve as a convenient model to study the activity of biotoxins consistently supplying with significant insights into the basic mechanisms of virus-host cell interactions and toxin entry into eukaryotic target cells. K1 and K2 toxins are active at the cell wall, leading to the disruption of the plasma membrane and subsequent cell death by ion...

  6. Three-Dimensional Analysis of Budding Sites and Released Virus Suggests a Revised Model for HIV-1 Morphogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Carlson, L.; Simon, M.; Briggs, J. A. G.; Glass, B.; Riches, J. D.; Johnson, M. C.; Muller, B.; Grunewald, K.; Krausslich, H.-G.

    2008-12-11

    Current models of HIV-1 morphogenesis hold that newly synthesized viral Gag polyproteins traffic to and assemble at the cell membrane into spherical protein shells. The resulting late-budding structure is thought to be released by the cellular ESCRT machinery severing the membrane tether connecting it to the producer cell. Using electron tomography and scanning transmission electron microscopy, we find that virions have a morphology and composition distinct from late-budding sites. Gag is arranged as a continuous but incomplete sphere in the released virion. In contrast, late-budding sites lacking functional ESCRT exhibited a nearly closed Gag sphere. The results lead us to propose that budding is initiated by Gag assembly, but is completed in an ESCRT-dependent manner before the Gag sphere is complete. This suggests that ESCRT functions early in HIV-1 release - akin to its role in vesicle formation - and is not restricted to severing the thin membrane tether.

  7. Genetic characterization of pathogenic fluorescent pseudomonads isolated from necrotic cherry and plum buds in Serbia

    Directory of Open Access Journals (Sweden)

    Gavrilović Veljko

    2013-01-01

    Full Text Available During past few years a symptoms of plum and cherry bud necrosis were observed in some regions with significant cherry production in Serbia. Gram negative, fluorescent, oxidative bacterial strains were isolated from the margin of necrotic tissue. All investigated strains are levan and HR positive, while negative results are recorded in oxidase, pectinase and arginin dihydrolase tests (LOPAT+---+. Symptoms similar to those observed in natural infection were obtained after artificial inoculation of cherry leaf scares and dormant one year old cherry shoots. Investigated strains as well as reference strain of P. syringae pv. morsprunorum cause the superficial necrosis on artificially inoculated immature cherry fruits, but negative results were recorded in immature pear and lemon fruit tests as well as syringae leaves and bean pods. Gelatin and aesculin tests were negative and tyrosinase and tartrate were positive. Investigated strains isolated from necrotic cherry buds had identical REP-PCR pattern with reference strain of P. syringae pv. morsprunorum. On the basis of obtained results, it was concluded that this bacterium is causal agent of cherry trees bud necrosis in Serbia. [Projekat Ministarstva nauke Republike Srbije, br. 31018 i br. 173026

  8. Ultraviolet irradiation initiates ectopic foot formation in regenerating hydra and promotes budding

    Indian Academy of Sciences (India)

    Saroj S Ghaskadbi; Leena Shetye; Shashi Chiplonkar; Surendra Ghaskadbi

    2005-03-01

    We have studied the effects of ultraviolet-C (UVC) and Ultraviolet-B (UVB) on growth and pattern formation in Pelmatohydra oligactis. UVC brings about a significant increase in budding in intact hydra while UVB does not exhibit such an effect. Excessive budding could be a response for survival at wavelengths that damage biological tissues. If the head or base piece of a bisected hydra is irradiated and recombined with the unirradiated missing part, regeneration proceeds normally indicating that exposure of a body part with either an intact head or foot to UVC does not influence pattern formation. Most significantly, in the middle piece, but not in the head or the base piece of a trisected hydra, UVC leads to initiation of ectopic feet formation in almost one third of the cases. Thus, UV irradiation interferes with pattern formation in regenerating hydra, possibly by changing positional values, and promotes budding in intact hydra. This is the first report on induction of ectopic feet formation by UV in regenerating hydra and opens up the possibility of using UV irradiation as a tool to understand pattern formation in the enigmatic hydra.

  9. Roles of the three L-domains in β-retrovirus budding.

    Science.gov (United States)

    Narahara, Chisato; Yasuda, Jiro

    2015-09-01

    Retroviral Gag protein plays a critical role during the late stage of virus budding and possesses a so-called L-domain containing PT/SAP, PPxY, YxxL or FPIV motifs that are critical for efficient budding. Mason-Pfizer monkey virus (M-PMV) contains PSAP, PPPY, and YADL sequences in Gag. This study was performed to investigate the roles of these three L-domain-like sequences in virus replication in three different cell lines, 293T, COS-7 and HeLa cells. It was found that the PPxY motif plays an essential role in progeny virus production as a major L-domain in all three cell lines. The PSAP sequence was shown to function as an additional L-domain in HeLa cells and to promote efficient release of M-PMV; however, this sequence was dispensable for M-PMV production in 293T and COS-7 cells, suggesting that the role of the PSAP motif as an L-domain in M-PMV budding is cell type-dependent. Viruses possessing multiple L-domains appear to change the L-domain usage to replicate in various cells. On the other hand, the YADL motif was required for M-PMV production as a transport signal of Gag to the plasma membrane, but not as an L-domain. PMID:26190584

  10. The Malleable Nature of the Budding Yeast Nuclear Envelope: Flares, Fusion, and Fenestrations.

    Science.gov (United States)

    Meseroll, Rebecca A; Cohen-Fix, Orna

    2016-11-01

    In eukaryotes, the nuclear envelope (NE) physically separates nuclear components and activities from rest of the cell. The NE also provides rigidity to the nucleus and contributes to chromosome organization. At the same time, the NE is highly dynamic; it must change shape and rearrange its components during development and throughout the cell cycle, and its morphology can be altered in response to mutation and disease. Here we focus on the NE of budding yeast, Saccharomyces cerevisiae, which has several unique features: it remains intact throughout the cell cycle, expands symmetrically during interphase, elongates during mitosis and, expands asymmetrically during mitotic delay. Moreover, its NE is safely breached during mating and when large structures, such as nuclear pore complexes and the spindle pole body, are embedded into its double membrane. The budding yeast NE lacks lamins and yet the nucleus is capable of maintaining a spherical shape throughout interphase. Despite these eccentricities, studies of the budding yeast NE have uncovered interesting, and likely conserved, processes that contribute to NE dynamics. In particular, we discuss the processes that drive and enable NE expansion and the dramatic changes in the NE that lead to extensions and fenestrations. J. Cell. Physiol. 231: 2353-2360, 2016. Published 2016. This article is a U.S. Government work and is in the public domain in the USA. PMID:26909870

  11. Bud Rot Caused by Phytophthora palmivora: A Destructive Emerging Disease of Oil Palm.

    Science.gov (United States)

    Torres, G A; Sarria, G A; Martinez, G; Varon, F; Drenth, A; Guest, D I

    2016-04-01

    Oomycetes from the genus Phytophthora are among the most important plant pathogens in agriculture. Epidemics caused by P. infestans precipitated the great Irish famine and had a major impact on society and human history. In the tropics, P. palmivora is a pathogen of many plant species including cacao (Theobroma cacao), citrus (Citrus sp.), durian (Durio zibethines), jackfruit (Artrocarpus heterophyllus), rubber (Hevea brasiliensis), and several palm species including coconut (Cocos nucifera), and the African oil palm (Elaeis guineensis) as determined recently. The first localized epidemics of bud rot in oil palm in Colombia were reported in 1964. However, recent epidemics of bud rot have destroyed more than 70,000 ha of oil palm in the Western and Central oil palm growing regions of Colombia. The agricultural, social, and economic implications of these outbreaks have been significant in Colombia. Identification of the pathogen after 100 years of investigating the disease in the world enabled further understanding of infection, expression of a range of symptoms, and epidemiology of the disease. This review examines the identification of P. palmivora as the cause of bud rot in Colombia, its epidemiology, and discusses the importance of P. palmivora as a major threat to oil palm plantings globally.

  12. dackel acts in the ectoderm of the zebrafish pectoral fin bud to maintain AER signaling.

    Science.gov (United States)

    Grandel, H; Draper, B W; Schulte-Merker, S

    2000-10-01

    Classical embryological studies have implied the existence of an apical ectodermal maintenance factor (AEMF) that sustains signaling from the apical ectodermal ridge (AER) during vertebrate limb development. Recent evidence suggests that AEMF activity is composed of different signals involving both a sonic hedgehog (Shh) signal and a fibroblast growth factor 10 (Fgf10) signal from the mesenchyme. In this study we show that the product of the dackel (dak) gene is one of the components that acts in the epidermis of the zebrafish pectoral fin bud to maintain signaling from the apical fold, which is homologous to the AER of tetrapods. dak acts synergistically with Shh to induce fgf4 and fgf8 expression but independently of Shh in promoting apical fold morphogenesis. The failure of dak mutant fin buds to progress from the initial fin induction phase to the autonomous outgrowth phase causes loss of both AER and Shh activity, and subsequently results in a proximodistal truncation of the fin, similar to the result obtained by ridge ablation experiments in the chicken. Further analysis of the dak mutant phenotype indicates that the activity of the transcription factor engrailed 1 (En1) in the ventral non-ridge ectoderm also depends on a maintenance signal probably provided by the ridge. This result uncovers a new interaction between the AER and the dorsoventral organizer in the zebrafish pectoral fin bud.

  13. Apical constriction initiates new bud formation during monopodial branching of the embryonic chicken lung

    Science.gov (United States)

    Kim, Hye Young; Varner, Victor D.; Nelson, Celeste M.

    2013-01-01

    Branching morphogenesis sculpts the airway epithelium of the lung into a tree-like structure to conduct air and promote gas exchange after birth. In the avian lung, a series of buds emerges from the dorsal surface of the primary bronchus via monopodial branching to form the conducting airways; anatomically, these buds are similar to those formed by domain branching in the mammalian lung. Here, we show that monopodial branching is initiated by apical constriction of the airway epithelium, and not by differential cell proliferation, using computational modeling and quantitative imaging of embryonic chicken lung explants. Both filamentous actin and phosphorylated myosin light chain were enriched at the apical surface of the airway epithelium during monopodial branching. Consistently, inhibiting actomyosin contractility prevented apical constriction and blocked branch initiation. Although cell proliferation was enhanced along the dorsal and ventral aspects of the primary bronchus, especially before branch formation, inhibiting proliferation had no effect on the initiation of branches. To test whether the physical forces from apical constriction alone are sufficient to drive the formation of new buds, we constructed a nonlinear, three-dimensional finite element model of the airway epithelium and used it to simulate apical constriction and proliferation in the primary bronchus. Our results suggest that, consistent with the experimental results, apical constriction is sufficient to drive the early stages of monopodial branching whereas cell proliferation is dispensable. We propose that initial folding of the airway epithelium is driven primarily by apical constriction during monopodial branching of the avian lung. PMID:23824575

  14. Potential bud bank responses to apical meristem damage and environmental variables: matching or complementing axillary meristems?

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    Jitka Klimešová

    Full Text Available Soil nutrients, dormant axillary meristem availability, and competition can influence plant tolerance to damage. However, the role of potential bud banks (adventitious meristems initiated only after injury is not known. Examining Central European field populations of 22 species of short-lived monocarpic herbs exposed to various sources of damage, we hypothesized that: (1 with increasing injury severity, the number of axillary branches would decrease, due to axillary meristem limitation, whereas the number of adventitious shoots (typically induced by severe injury would increase; (2 favorable environmental conditions would allow intact plants to branch more, resulting in stronger axillary meristem limitation than in unfavorable conditions; and (3 consequently, adventitious sprouting would be better enabled in favorable than unfavorable conditions. We found strong support for the first hypothesis, only limited support for the second, and none for the third. Our results imply that whereas soil nutrients and competition marginally influence plant tolerance to damage, potential bud banks enable plants to overcome meristem limitation from severe damage, and therefore better tolerate it. All the significant effects were found in intraspecific comparisons, whereas interspecific differences were not found. Monocarpic plants with potential bud banks therefore represent a distinct strategy occupying a narrow environmental niche. The disturbance regime typical for this niche remains to be examined, as do the costs associated with the banks of adventitious and axillary reserve meristems.

  15. Potential bud bank responses to apical meristem damage and environmental variables: matching or complementing axillary meristems?

    Science.gov (United States)

    Klimešová, Jitka; Malíková, Lenka; Rosenthal, Jonathan; Šmilauer, Petr

    2014-01-01

    Soil nutrients, dormant axillary meristem availability, and competition can influence plant tolerance to damage. However, the role of potential bud banks (adventitious meristems initiated only after injury) is not known. Examining Central European field populations of 22 species of short-lived monocarpic herbs exposed to various sources of damage, we hypothesized that: (1) with increasing injury severity, the number of axillary branches would decrease, due to axillary meristem limitation, whereas the number of adventitious shoots (typically induced by severe injury) would increase; (2) favorable environmental conditions would allow intact plants to branch more, resulting in stronger axillary meristem limitation than in unfavorable conditions; and (3) consequently, adventitious sprouting would be better enabled in favorable than unfavorable conditions. We found strong support for the first hypothesis, only limited support for the second, and none for the third. Our results imply that whereas soil nutrients and competition marginally influence plant tolerance to damage, potential bud banks enable plants to overcome meristem limitation from severe damage, and therefore better tolerate it. All the significant effects were found in intraspecific comparisons, whereas interspecific differences were not found. Monocarpic plants with potential bud banks therefore represent a distinct strategy occupying a narrow environmental niche. The disturbance regime typical for this niche remains to be examined, as do the costs associated with the banks of adventitious and axillary reserve meristems. PMID:24516587

  16. A mathematical model for the induction of the mammalian ureteric bud.

    Science.gov (United States)

    Lawson, Brodie A J; Flegg, Mark B

    2016-04-01

    Congenital abnormalities of the kidney and urinary tract collectively form the most common type of prenatally diagnosed malformations. Whilst many of the crucial genes that direct the kidney developmental program are known, the mechanisms by which kidney organogenesis is achieved is still largely unclear. In this paper, we propose a mathematical model for the localisation of the ureteric bud, the precursor to the ureter and collecting duct system of the kidney. The mathematical model presented fundamentally implicates Schnakenberg-like ligand-receptor Turing patterning as the mechanism by which the ureteric bud is localised on the Wolfian duct as proposed by Menshykaul and Iber (2013). This model explores the specific roles of regulatory proteins GREM1 and BMP as well as the domain properties of GDNF production. Our model demonstrates that this proposed pattern formation mechanism is capable of naturally predicting the phenotypical outcomes of many genetic experiments from the literature. Furthermore, we conclude that whilst BMP inhibits GDNF away from the budding site and GREM1 permits GDNF to signal, GREM1 also stabilises the effect of BMP on GDNF signalling from fluctuations in BMP sensitivity but not signal strength. PMID:26801874

  17. Bud Rot Caused by Phytophthora palmivora: A Destructive Emerging Disease of Oil Palm.

    Science.gov (United States)

    Torres, G A; Sarria, G A; Martinez, G; Varon, F; Drenth, A; Guest, D I

    2016-04-01

    Oomycetes from the genus Phytophthora are among the most important plant pathogens in agriculture. Epidemics caused by P. infestans precipitated the great Irish famine and had a major impact on society and human history. In the tropics, P. palmivora is a pathogen of many plant species including cacao (Theobroma cacao), citrus (Citrus sp.), durian (Durio zibethines), jackfruit (Artrocarpus heterophyllus), rubber (Hevea brasiliensis), and several palm species including coconut (Cocos nucifera), and the African oil palm (Elaeis guineensis) as determined recently. The first localized epidemics of bud rot in oil palm in Colombia were reported in 1964. However, recent epidemics of bud rot have destroyed more than 70,000 ha of oil palm in the Western and Central oil palm growing regions of Colombia. The agricultural, social, and economic implications of these outbreaks have been significant in Colombia. Identification of the pathogen after 100 years of investigating the disease in the world enabled further understanding of infection, expression of a range of symptoms, and epidemiology of the disease. This review examines the identification of P. palmivora as the cause of bud rot in Colombia, its epidemiology, and discusses the importance of P. palmivora as a major threat to oil palm plantings globally. PMID:26714102

  18. Bud burst and flowering phenology in a mixed oak forest from Eastern Romania

    Directory of Open Access Journals (Sweden)

    Ecaterina Nicoleta Chesnoiu

    2009-12-01

    Full Text Available Bud burst and flowering phenology have been observed in year 2008 in a natural white oak species complex situated in eastern Romania. A total of 300 mature individuals was mapped and identified based on leaf morphology. The community consists of four oak species: Quercus pedunculiflora, Q. robur, Q. pubescens and Q. petraea. A set of 28 individuals could not be unambiguously classified to one or another species. Data on bud burst showed a normal distribution and the differences among species were small. The "very late" flushing was recorded on 15th of April, three weeks later when compared to early flushing individuals. The time period between the bud burst and the complete development of leaves was nearly the same in all oak species, varying on average, between 18.4 and 20.6 days. The spatial distribution of phenological groups within the complex appears to be non-randomly, because in many parts of the study plot exist groups in which most of the trees belong to the same phenological category. Our results indicate an overlap in flowering time for all oak species which occur in the area. The data support the hypothesis that interspecific gene flow is possible between closely related oak species.

  19. SUBSTRATES UTILIZATION TO ASSESS ROOTEDNESS CAPACITY AND VIABILITY BUDS AT SOME GRAPE VARIETIES

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    Gheorghe Cristian Popescu

    2013-12-01

    Full Text Available The cultivated grapevine (Vitis vinifera L. is a fruit crop of enormous economic importance with over eight million hectares planted in vineyards worldwide. Table grapes and wines represent a considerable share of the economy in many grape and wine-producing countries. During the dormant, due to low temperatures and how to prepare grape for entrance in winter time, wood annual increases and buds may be adversely affected. The way how the vines passed by dormant period can affect the buds and wood viability and rooting ability of vine cuttings. In this study were tested on different culture substrates vine cuttings belonging to a noble variety and a hybrid vines: Merlot and Isabella. Noble grapes are a term used to describe the international variety of grapes that are most recognizable for the top quality wine they produce. In this paper was determinate total dry matter of vine cuttings, humidity of biological material, vine cuttings rooting capacity and viability status buds cuttings placed on three nutritional substrates.

  20. Nutrition induced pleomorphism and budding mode of reproduction in Deinococcus radiodurans

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    Toleti Rao S

    2009-07-01

    Full Text Available Abstract Background Morphological adaptation is an important biological function of a microorganism to cope with its environment. Pleomorphism (to exist in a number of morphological forms took centre stage in many discussions wherein a bacterium exhibits morphological transition and altered mode of reproduction in response to an environmental condition. Findings To strengthen the concept on pleomorphism in bacteria, we report here different cell morphologies of Deinococcus radiodurans in response to variation in nutrient concentration. From our studies we attempt primary evidence towards the presence of significant population of monomer cells of D. radiodurans in specific culture condition. In this report we also illustrate with scanning electron micrographs an unusual budding mode of reproduction in D. radiodurans which was not reported till date for this group of bacteria. Conclusion In a holistic view the study reflects on bacterial shape (morphotypes and the physiological adaptation to a particular nutrient environment. The discovery of budding mode of reproduction in Deinococcus will be of interest to microbiologists. It can serve as a model system to understand the mechanism of budding process at molecular level.

  1. cDNA sequence quality data - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available Budding yeast cDNA sequencing project cDNA sequence quality data Data detail Data name cDNA sequence quality... data Description of data contents Phred's quality score. PHD format, one file to a single cDNA data, and co...ription Download License Update History of This Database Site Policy | Contact Us cDNA sequence quality data - Budding yeast cDNA sequencing project | LSDB Archive ...

  2. Genetic analysis reveals an unexpected role of BMP7 in initiation of ureteric bud outgrowth in mouse embryos.

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    Alexandre Gonçalves

    Full Text Available BACKGROUND: Genetic analysis in the mouse revealed that GREMLIN1 (GREM1-mediated antagonism of BMP4 is essential for ureteric epithelial branching as the disruption of ureteric bud outgrowth and renal agenesis in Grem1-deficient embryos is restored by additional inactivation of one Bmp4 allele. Another BMP ligand, BMP7, was shown to control the proliferative expansion of nephrogenic progenitors and its requirement for nephrogenesis can be genetically substituted by Bmp4. Therefore, we investigated whether BMP7 in turn also participates in inhibiting ureteric bud outgrowth during the initiation of metanephric kidney development. METHODOLOGY/PRINCIPAL FINDINGS: Genetic inactivation of one Bmp7 allele in Grem1-deficient mouse embryos does not alleviate the bilateral renal agenesis, while complete inactivation of Bmp7 restores ureteric bud outgrowth and branching. In mouse embryos lacking both Grem1 and Bmp7, GDNF/WNT11 feedback signaling and the expression of the Etv4 target gene, which regulates formation of the invading ureteric bud tip, are restored. In contrast to the restoration of ureteric bud outgrowth and branching, nephrogenesis remains aberrant as revealed by the premature loss of Six2 expressing nephrogenic progenitor cells. Therefore, very few nephrons develop in kidneys lacking both Grem1 and Bmp7 and the resulting dysplastic phenotype is indistinguishable from the one of Bmp7-deficient mouse embryos. CONCLUSIONS/SIGNIFICANCE: Our study reveals an unexpected inhibitory role of BMP7 during the onset of ureteric bud outgrowth. As BMP4, BMP7 and GREM1 are expressed in distinct mesenchymal and epithelial domains, the localized antagonistic interactions of GREM1 with BMPs could restrict and guide ureteric bud outgrowth and branching. The robustness and likely significant redundancy of the underlying signaling system is evidenced by the fact that global reduction of Bmp4 or inactivation of Bmp7 are both able to restore ureteric bud outgrowth

  3. The conserved Bud20 zinc finger protein is a new component of the ribosomal 60S subunit export machinery.

    Science.gov (United States)

    Bassler, Jochen; Klein, Isabella; Schmidt, Claudia; Kallas, Martina; Thomson, Emma; Wagner, Maria Anna; Bradatsch, Bettina; Rechberger, Gerald; Strohmaier, Heimo; Hurt, Ed; Bergler, Helmut

    2012-12-01

    The nuclear export of the preribosomal 60S (pre-60S) subunit is coordinated with late steps in ribosome assembly. Here, we show that Bud20, a conserved C(2)H(2)-type zinc finger protein, is an unrecognized shuttling factor required for the efficient export of pre-60S subunits. Bud20 associates with late pre-60S particles in the nucleoplasm and accompanies them into the cytoplasm, where it is released through the action of the Drg1 AAA-ATPase. Cytoplasmic Bud20 is then reimported via a Kap123-dependent pathway. The deletion of Bud20 induces a strong pre-60S export defect and causes synthetic lethality when combined with mutant alleles of known pre-60S subunit export factors. The function of Bud20 in ribosome export depends on a short conserved N-terminal sequence, as we observed that mutations or the deletion of this motif impaired 60S subunit export and generated the genetic link to other pre-60S export factors. We suggest that the shuttling Bud20 is recruited to the nascent 60S subunit via its central zinc finger rRNA binding domain to facilitate the subsequent nuclear export of the preribosome employing its N-terminal extension.

  4. Plant regeneration and floral bud formation from intact floral parts of African violet (Saintpaulia ionantha H. Wendl.) cultured in vitro.

    Science.gov (United States)

    Daud, N; Taha, R M

    2008-04-01

    Intact immature flower buds of African violet (Saintpaulia ionantha H. Wendl.) were used as explant sources for in vitro studies. The effect of exogenous hormones, NAA and BAP on the indirect organogenesis of this species was observed. Callus was formed on the cut end (base) of pedicels of floral buds where they were in contact with the medium. When maintained on the same medium, callus was differentiated into adventitious shoots after 10 weeks in culture. MS media supplemented with 2.0 mg L(-1) NAA and 1.0 mg L(-1) BAP gave the highest number of sterile or vegetative floral buds from the surface of callus of the explants, but these buds failed to develop further. The floral buds were expanded as abnormal flowers. The floral structures were smaller in size compared to intact flowers. Petals (corolla) were white to purple in colour but did not form any reproductive organs, i.e., stamens or pistils. All sterile or vegetative floral buds and abnormal flowers survived for 3 months in culture but failed to reach anthesis. PMID:18810979

  5. The effect of the times and the budding methods on the quality of young trees and the nursery efficiency of cherry trees cv. 'Łutówka'

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    Piotr Baryła

    2012-12-01

    Full Text Available The studies concerning the effect of the times and the methods of budding on the growth of young cherry trees were conducted in the years 1997-2000 at Felin Experimental Farm of Lublin Agricultural University. The objects of investigations were the young cherry trees obtained as a result of budding of mahaleb cherry (Prunus mahaleb L. and sweet cherry (Prunus avium L. seedlings in the way by the chip budding-15th July and T-budding-on the 15th July and the 1st September. The used terms and budding methods did not affect the bud taking and the quality of cherry trees during three years studies. Chip budding of the sweet cherry on the 15th July was the most effective way of this seedling budding. Late budding-on the 1st September-did not change the efficiency of the nursery only in case of mahaleb cherry. The highest number-33 000 of the young trees, average per 1 ha was got as a result of the chip and "T" mahaleb cherry budding on the 1st September.

  6. Change in Auxin and Cytokinin Levels Coincides with Altered Expression of Branching Genes during Axillary Bud Outgrowth in Chrysanthemum.

    Science.gov (United States)

    Dierck, Robrecht; De Keyser, Ellen; De Riek, Jan; Dhooghe, Emmy; Van Huylenbroeck, Johan; Prinsen, Els; Van Der Straeten, Dominique

    2016-01-01

    In the production and breeding of Chrysanthemum sp., shoot branching is an important quality aspect as the outgrowth of axillary buds determines the final plant shape. Bud outgrowth is mainly controlled by apical dominance and the crosstalk between the plant hormones auxin, cytokinin and strigolactone. In this work the hormonal and genetic regulation of axillary bud outgrowth was studied in two differently branching cut flower Chrysanthemum morifolium (Ramat) genotypes. C17 is a split-type which forms an inflorescence meristem after a certain vegetative period, while C18 remains vegetative under long day conditions. Plant growth of both genotypes was monitored during 5 subsequent weeks starting one week before flower initiation occurred in C17. Axillary bud outgrowth was measured weekly and samples of shoot apex, stem and axillary buds were taken during the first two weeks. We combined auxin and cytokinin measurements by UPLC-MS/MS with RT-qPCR expression analysis of genes involved in shoot branching regulation pathways in chrysanthemum. These included bud development genes (CmBRC1, CmDRM1, CmSTM, CmLsL), auxin pathway genes (CmPIN1, CmTIR3, CmTIR1, CmAXR1, CmAXR6, CmAXR2, CmIAA16, CmIAA12), cytokinin pathway genes (CmIPT3, CmHK3, CmRR1) and strigolactone genes (CmMAX1 and CmMAX2). Genotype C17 showed a release from apical dominance after floral transition coinciding with decreased auxin and increased cytokinin levels in the subapical axillary buds. As opposed to C17, C18 maintained strong apical dominance with vegetative growth throughout the experiment. Here high auxin levels and decreasing cytokinin levels in axillary buds and stem were measured. A differential expression of several branching genes accompanied the different hormonal change and bud outgrowth in C17 and C18. This was clear for the strigolactone biosynthesis gene CmMAX1, the transcription factor CmBRC1 and the dormancy associated gene CmDRM1, that all showed a decreased expression in C17 at floral

  7. Ubiquitin-regulated nuclear-cytoplasmic trafficking of the Nipah virus matrix protein is important for viral budding.

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    Yao E Wang

    Full Text Available Paramyxoviruses are known to replicate in the cytoplasm and bud from the plasma membrane. Matrix is the major structural protein in paramyxoviruses that mediates viral assembly and budding. Curiously, the matrix proteins of a few paramyxoviruses have been found in the nucleus, although the biological function associated with this nuclear localization remains obscure. We report here that the nuclear-cytoplasmic trafficking of the Nipah virus matrix (NiV-M protein and associated post-translational modification play a critical role in matrix-mediated virus budding. Nipah virus (NiV is a highly pathogenic emerging paramyxovirus that causes fatal encephalitis in humans, and is classified as a Biosafety Level 4 (BSL4 pathogen. During live NiV infection, NiV-M was first detected in the nucleus at early stages of infection before subsequent localization to the cytoplasm and the plasma membrane. Mutations in the putative bipartite nuclear localization signal (NLS and the leucine-rich nuclear export signal (NES found in NiV-M impaired its nuclear-cytoplasmic trafficking and also abolished NiV-M budding. A highly conserved lysine residue in the NLS served dual functions: its positive charge was important for mediating nuclear import, and it was also a potential site for monoubiquitination which regulates nuclear export of the protein. Concordantly, overexpression of ubiquitin enhanced NiV-M budding whereas depletion of free ubiquitin in the cell (via proteasome inhibitors resulted in nuclear retention of NiV-M and blocked viral budding. Live Nipah virus budding was exquisitely sensitive to proteasome inhibitors: bortezomib, an FDA-approved proteasome inhibitor for treating multiple myeloma, reduced viral titers with an IC(50 of 2.7 nM, which is 100-fold less than the peak plasma concentration that can be achieved in humans. This opens up the possibility of using an "off-the-shelf" therapeutic against acute NiV infection.

  8. Shoot bending promotes flower bud formation by miRNA-mediated regulation in apple (Malus domestica Borkh.).

    Science.gov (United States)

    Xing, Libo; Zhang, Dong; Zhao, Caiping; Li, Youmei; Ma, Juanjuan; An, Na; Han, Mingyu

    2016-02-01

    Flower induction in apple (Malus domestica Borkh.) trees plays an important life cycle role, but young trees produce fewer and inferior quality flower buds. Therefore, shoot bending has become an important cultural practice, significantly promoting the capacity to develop more flower buds during the growing seasons. Additionally, microRNAs (miRNAs) play essential roles in plant growth, flower induction and stress responses. In this study, we identified miRNAs potentially involved in the regulation of bud growth, and flower induction and development, as well as in the response to shoot bending. Of the 195 miRNAs identified, 137 were novel miRNAs. The miRNA expression profiles revealed that the expression levels of 68 and 27 known miRNAs were down-regulated and up-regulated, respectively, in response to shoot bending, and that the 31 differentially expressed novel miRNAs between them formed five major clusters. Additionally, a complex regulatory network associated with auxin, cytokinin, abscisic acid (ABA) and gibberellic acid (GA) plays important roles in cell division, bud growth and flower induction, in which related miRNAs and targets mediated regulation. Among them, miR396, 160, 393, and their targets associated with AUX, miR159, 319, 164, and their targets associated with ABA and GA, and flowering-related miRNAs and genes, regulate bud growth and flower bud formation in response to shoot bending. Meanwhile, the flowering genes had significantly higher expression levels during shoot bending, suggesting that they are involved in this regulatory process. This study provides a framework for the future analysis of miRNAs associated with multiple hormones and their roles in the regulation of bud growth, and flower induction and formation in response to shoot bending in apple trees.

  9. Disruption of microtubules uncouples budding and nuclear division in Toxoplasma gondii.

    Science.gov (United States)

    Morrissette, Naomi S; Sibley, L David

    2002-03-01

    The tachyzoite stage of the protozoan parasite Toxoplasma gondii has two populations of microtubules: spindle microtubules and subpellicular microtubules. To determine how these two microtubule populations are regulated, we investigated microtubule behavior during the cell cycle following treatment with microtubule-disrupting drugs. Previous work had established that the microtubule populations are individually nucleated by two distinct microtubule-organizing centers (MTOCs): the apical polar ring for the subpellicular microtubules and spindle pole plaques/centrioles for the spindle microtubules. When replicating tachyzoites were treated with 0.5 microM oryzalin or 1.0 mM colchicine they retained the capacity to form a spindle and undergo nuclear division. Although these parasites could complete budding, they lost the bulk of their subpellicular microtubules and the ability to reinvade host cells. Both nascent spindle and subpellicular microtubules were disrupted in 2.5 microM oryzalin or 5.0 mM colchicine. Under these conditions, parasites grew in size and replicated their genome but were incapable of nuclear division. After removal from 0.5 microM oryzalin, Toxoplasma tachyzoites were able to restore normal subpellicular microtubules and a fully invasive phenotype. When oryzalin was removed from Toxoplasma tachyzoites treated with 2.5 microM drug, the parasites attempted to bud as crescent-shaped tachyzoites. Because the polyploid nuclear mass could not be correctly segregated, many daughter parasites lacked nuclei altogether although budding and scission from the maternal mass was able to be completed. Multiple MTOCs permit Toxoplasma tachyzoites to control nuclear division independently from cell polarity and cytokinesis. This unusual situation grants greater cell cycle flexibility to these parasites but abolishes the checks for coregulation of nuclear division and cytokinesis found in other eukaryotes. PMID:11870220

  10. Genes in the ureteric budding pathway: association study on vesico-ureteral reflux patients.

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    Albertien M van Eerde

    Full Text Available Vesico-ureteral reflux (VUR is the retrograde passage of urine from the bladder to the urinary tract and causes 8.5% of end-stage renal disease in children. It is a complex genetic developmental disorder, in which ectopic embryonal ureteric budding is implicated in the pathogenesis. VUR is part of the spectrum of Congenital Anomalies of the Kidney and Urinary Tract (CAKUT. We performed an extensive association study for primary VUR using a two-stage, case-control design, investigating 44 candidate genes in the ureteric budding pathway in 409 Dutch VUR patients. The 44 genes were selected from the literature and a set of 567 single nucleotide polymorphisms (SNPs capturing their genetic variation was genotyped in 207 cases and 554 controls. The 14 SNPs with p<0.005 were included in a follow-up study in 202 cases and 892 controls. Of the total cohort, ~50% showed a clear-cut primary VUR phenotype and ~25% had both a duplex collecting system and VUR. We also looked for association in these two extreme phenotype groups. None of the SNPs reached a significant p-value. Common genetic variants in four genes (GREM1, EYA1, ROBO2 and UPK3A show a trend towards association with the development of primary VUR (GREM1, EYA1, ROBO2 or duplex collecting system (EYA1 and UPK3A. SNPs in three genes (TGFB1, GNB3 and VEGFA have been shown to be associated with VUR in other populations. Only the result of rs1800469 in TGFB1 hinted at association in our study. This is the first extensive study of common variants in the genes of the ureteric budding pathway and the genetic susceptibility to primary VUR.

  11. Screening the budding yeast genome reveals unique factors affecting K2 toxin susceptibility.

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    Elena Servienė

    Full Text Available BACKGROUND: Understanding how biotoxins kill cells is of prime importance in biomedicine and the food industry. The budding yeast (S. cerevisiae killers serve as a convenient model to study the activity of biotoxins consistently supplying with significant insights into the basic mechanisms of virus-host cell interactions and toxin entry into eukaryotic target cells. K1 and K2 toxins are active at the cell wall, leading to the disruption of the plasma membrane and subsequent cell death by ion leakage. K28 toxin is active in the cell nucleus, blocking DNA synthesis and cell cycle progression, thereby triggering apoptosis. Genome-wide screens in the budding yeast S. cerevisiae identified several hundred effectors of K1 and K28 toxins. Surprisingly, no such screen had been performed for K2 toxin, the most frequent killer toxin among industrial budding yeasts. PRINCIPAL FINDINGS: We conducted several concurrent genome-wide screens in S. cerevisiae and identified 332 novel K2 toxin effectors. The effectors involved in K2 resistance and hypersensitivity largely map in distinct cellular pathways, including cell wall and plasma membrane structure/biogenesis and mitochondrial function for K2 resistance, and cell wall stress signaling and ion/pH homeostasis for K2 hypersensitivity. 70% of K2 effectors are different from those involved in K1 or K28 susceptibility. SIGNIFICANCE: Our work demonstrates that despite the fact that K1 and K2 toxins share some aspects of their killing strategies, they largely rely on different sets of effectors. Since the vast majority of the host factors identified here is exclusively active towards K2, we conclude that cells have acquired a specific K2 toxin effectors set. Our work thus indicates that K1 and K2 have elaborated different biological pathways and provides a first step towards the detailed characterization of K2 mode of action.

  12. The Relation Between Endogenous Hormones and Late-Germination in Buds of Avrolles Apple

    Institute of Scientific and Technical Information of China (English)

    QIN Dong; WANG Jin-zheng; GUO Jian-min; ZHAI Heng

    2009-01-01

    In order to provide the physiological bases for selecting late-germination cultivars that can avoid late frost damage,the very late-germination variety Avrolles (Malus domestica) was used to study the relation between the dynamic changes and balance of endogenous hormones and germination time.The concentrations of endogenous GA3,ABA,IAA,and ZR were determined in buds of Avrolles and Judeline (Malus domestica) from dormancy releasing to germination by capillary electrophoresis.The dynamic changes of endogenous hormones concentration in buds of Avrolles and Judeline were similar; but the magnitude and time of the change were significantly different between the two varieties,especially for GA3.GA3 concentration increased with dormancy releasing,then decreased,and increased again before germination in the two varieties.GA3 concentration in Avrolles was 1.72 times that in Judeline at the first peak,the gap increased to 2.22 times at germination.ZR concentration exhibited a continuous increase trend,but it decreased sharply before germination.ZR accumulation in Avrolles took 36 days longer than in Judeline,the peak value was 44% higher than in Judeline.Before germination,ZR concentration in Avrolles was 2.12 times that in Judeline.The differences between IAA and ABA concentration were relatively small in the two varieties,while the ratios of GA3/ABA and (GA3 + IAA + ZR)/ABA in Avrolles were 2.08 and 1.58 times those in Judeline,respectively.The germination of apple bud was regulated by the endogenous hormones.For the late-germination apple Avrolles,its germination requires higher concentration of GA3 and ZR,which leads to the high ratios of GA3/ABA and (GA3 + IAA + ZR)/ABA.

  13. Immunocytochemical analysis of P2X2 in rat circumvallate taste buds

    Directory of Open Access Journals (Sweden)

    Yang Ruibiao

    2012-05-01

    Full Text Available Abstract Background Our laboratory has shown that classical synapses and synaptic proteins are associated with Type III cells. Yet it is generally accepted that Type II cells transduce bitter, sweet and umami stimuli. No classical synapses, however, have been found associated with Type II cells. Recent studies indicate that the ionotropic purinergic receptors P2X2/P2X3 are present in rodent taste buds. Taste nerve processes express the ionotropic purinergic receptors (P2X2/P2X3. P2X2/P2X3Dbl−/− mice are not responsive to sweet, umami and bitter stimuli, and it has been proposed that ATP acts as a neurotransmitter in taste buds. The goal of the present study is to learn more about the nature of purinergic contacts in rat circumvallate taste buds by examining immunoreactivity to antisera directed against the purinergic receptor P2X2. Results P2X2-like immunoreactivity is present in intragemmal nerve processes in rat circumvallate taste buds. Intense immunoreactivity can also be seen in the subgemmal nerve plexuses located below the basal lamina. The P2X2 immunoreactive nerve processes also display syntaxin-1-LIR. The immunoreactive nerves are in close contact with the IP3R3-LIR Type II cells and syntaxin-1-LIR and/or 5-HT-LIR Type III cells. Taste cell synapses are observed only from Type III taste cells onto P2X2-LIR nerve processes. Unusually large, “atypical” mitochondria in the Type II taste cells are found only at close appositions with P2X2-LIR nerve processes. P2X2 immunogold particles are concentrated at the membranes of nerve processes at close appositions with taste cells. Conclusions Based on our immunofluorescence and immunoelectron microscopical studies we believe that both perigemmal and most all intragemmal nerve processes display P2X2-LIR. Moreover, colloidal gold immunoelectron microscopy indicates that P2X2-LIR in nerve processes is concentrated at sites of close apposition with Type II cells. This supports the hypothesis

  14. In-vitro radiosensitivity and rapid propagation of potato through axillary bud culture

    International Nuclear Information System (INIS)

    The axillary buds of two varieties of potato, Cardinal and Ultimus, were cultured on MS medium supplemented with different concentrations and combinations of cytokinins, auxin and GA3 for their rapid propagation. The results indicated that combination: kinetin 0.5 + NAA 0.1 + GA3 0.5 mg/l proved to be the best for regeneration of maximum number of plantlets. Moreover, the mortality rates at 30 Gray dose in the varieties Cardinal and Ultimus were 41.6 and 58.3 percent respectively and hence this dose was considered to be the optimum for further mutation experiments in potato

  15. License - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available Budding yeast cDNA sequencing project License to Use This Database Last updated : 2010/02/15 You may use this database...scribed below. The Standard License specifies the license terms regarding the use of this database... and the requirements you must follow in using this database. The Additional License specif...icense. Standard License The Standard License for this database is the license sp...ecified in the Creative Commons Attribution-Share Alike 2.1 Japan . If you use data from this database, plea

  16. Soluble proteins and polyphenoloxidase activity in bud flowers, flowers and leaves of cold stored lisianthus

    DEFF Research Database (Denmark)

    Cavasini, R.; Nunes, K.N.M.; Favero, B.T.;

    This study evaluated the activity of the enzyme polyphenol oxidase (PPO) and the content of soluble protein present in lisianthus bud flowers, flowers and leaves in room temperature (24±2°C) and pre-exposure cold chamber at 9±2°C for 24 h, in order to examine a possible correlation between...... these parameters and postharvest longevity of lisianthus flowers. After treatments, flowers were kept in pots with water, stored at room temperature and evaluated every three days until the end of their decorative life for biochemical analyzes. During the experimental period the enzymatic activity increased...

  17. Pickling process of capers (Capparis spp. flower buds

    Directory of Open Access Journals (Sweden)

    Özcan, Musa

    1999-04-01

    Full Text Available Middle sized (8 < x < 13 mm buds of Capparis spinosa var. spinosa and C. ovata var. canescens from June in brines containing 5,10,15 and 20% salt and from August in brines of 15% salt, and three different size (x < 8 mm, 8 < x < 13 mm, x > 13 mm buds of C. . ovata var. canescens from June in brines of 15% salt were pickled for two months fermentation. Some chemical and microbiological analyses were done in brines during fermentation. Most suitable salt concentration for lactic acid bacteria (LAB activity were 5% and partly 10%. Acidity, LAB activity, sedimentation and hardness were reduced by increasing bud size in C. ovata. Small buds of C. ovata for pickling product had advantage for colour and flavour, however, more sediment and partly softening showed disadvantage. For both species, pickling time was determined as 40 to 50 days in regard of end-product flavour and odour, brine acidity and pH, and LAB activity.

    Se encurtieron durante dos meses botones florales de tamaño medio (8 < x < 13 mm de Capparis spinosa var. spinosa y C. ovata var. canescens, los recolectados en Junio en salmueras conteniendo 5, 10, 15 y 20% de sal, y los de Agosto en salmueras de 15% de sal; y tres tamaños diferentes (x < 8 mm, 8 < x < 13 mm, X > 13 mm de C. ovata var. canescens de Junio en salmueras de 15% de sal. Se realizaron algunos análisis químicos y microbiológicos durante la fermentación. Las concentraciones de sal más adecuadas para la actividad de las bacterias del ácido láctico (LAB fueron 5% y parcialmente 10%. Acidez, actividad de LAB, sedimentación y firmeza (hardness se redujeron al incrementar el tamaño de las alcaparras de C. ovata. Los tamaños pequeños de C. ovata presentaron en el producto encurtido ventajas en color y sabor, pero desventajas por más sedimento y ablandamiento parcial. El tiempo de encurtido para ambas

  18. Sponge budding is a spatiotemporal morphological patterning process: Insights from synchrotron radiation-based x-ray microtomography into the asexual reproduction of Tethya wilhelma

    Directory of Open Access Journals (Sweden)

    Nickel Michael

    2009-09-01

    Full Text Available Abstract Background Primary agametic-asexual reproduction mechanisms such as budding and fission are present in all non-bilaterian and many bilaterian animal taxa and are likely to be metazoan ground pattern characters. Cnidarians display highly organized and regulated budding processes. In contrast, budding in poriferans was thought to be less specific and related to the general ability of this group to reorganize their tissues. Here we test the hypothesis of morphological pattern formation during sponge budding. Results We investigated the budding process in Tethya wilhelma (Demospongiae by applying 3D morphometrics to high resolution synchrotron radiation-based x-ray microtomography (SR-μCT image data. We followed the morphogenesis of characteristic body structures and identified distinct morphological states which indeed reveal characteristic spatiotemporal morphological patterns in sponge bud development. We discovered the distribution of skeletal elements, canal system and sponge tissue to be based on a sequential series of distinct morphological states. Based on morphometric data we defined four typical bud stages. Once they have reached the final stage buds are released as fully functional juvenile sponges which are morphologically and functionally equivalent to adult specimens. Conclusion Our results demonstrate that budding in demosponges is considerably more highly organized and regulated than previously assumed. Morphological pattern formation in asexual reproduction with underlying genetic regulation seems to have evolved early in metazoans and was likely part of the developmental program of the last common ancestor of all Metazoa (LCAM.

  19. Differences between the Bud End and Stem End of Potatoes in Dry Matter Content, Starch Granule Size, and Carbohydrate Metabolic Gene Expression at the Growing and Sprouting Stages.

    Science.gov (United States)

    Liu, Bailin; Zhang, Guodong; Murphy, Agnes; De Koeyer, David; Tai, Helen; Bizimungu, Benoit; Si, Huaijun; Li, Xiu-Qing

    2016-02-10

    Potatoes usually have the tuber bud end dominance in growth during tuber bulking and in tuber sprouting, likely using carbohydrates from the tuber stem end. We hypothesized that the tuber bud end and tuber stem end coordination in carbohydrate metabolism gene expression is different between the bulking dominance and sprouting dominance of the tuber bud end. After comparing the growing tubers at harvest from a green vine and the stage that sprouts just started to emerge after storage of tubers at room temperature, we found the following: (1) Dry matter content was higher in the tuber stem end than the tuber bud end at both stages. (2) The starch granule size was larger in the tuber bud end than in the tuber stem end. (3) The tuber bud end had higher gene expression for starch synthesis but a lower gene expression of sucrose transporters than the tuber stem end during tuber growing. (4) The tuber stem end at the sprouting stage showed more active gene expression in both starch degradation and resynthesis, suggesting more active export of carbohydrates, than the tuber bud end. The results indicate that the starch accumulation mechanism in the tuber bud end was different between field growing and post-harvest sprouting tubers and that tubers already increased dry matter and average starch granule sizes in the tuber bud end prior to the rapid growth of sprouts. PMID:26760673

  20. Identification of differentially expressed genes associated with bud dormancy release in tree peony (Paeonia suffruticosa) by suppression subtractive hybridization

    Institute of Scientific and Technical Information of China (English)

    HUANG Xin; ZHENG Guo-sheng; DAI Si-lan; GAI Shu-peng

    2008-01-01

    A subtractive cDNA library was developed to study genes associated with bud dormancy release in tree peonies. In order to identify genes that are highly expressed in buds released from dormancy, 588 clones were examined by differential screening. Of these, 185 clones were selected to be sequenced. A total of 37 unique sequences were obtained of which only 31 sequences have matches in the NCBI database or the Arabidopsis thaliana protein database. Semi-quantitative RT-PCR was used to confirm further the expression profiles for 12 transcripts identified within the subtraetive cDNA library. Gene ontology analyses indicated that many of the different genes identified have unknown or hypothetical functions while it is speculated that other genes play different molecular roles. In our study, genes involved in bud dormancy release were growth-related or stress-responsive, while low-temperature-induced ribosomal proteins may also play a role in bud dormancy release. Our results provide interesting information for further understanding of the molecular mechanism of bud dormancy release in tree peonies.

  1. Bimolecular Complementation to Visualize Filovirus VP40-Host Complexes in Live Mammalian Cells: Toward the Identification of Budding Inhibitors

    Directory of Open Access Journals (Sweden)

    Yuliang Liu

    2011-01-01

    Full Text Available Virus-host interactions play key roles in promoting efficient egress of many RNA viruses, including Ebola virus (EBOV or “e” and Marburg virus (MARV or “m”. Late- (L- domains conserved in viral matrix proteins recruit specific host proteins, such as Tsg101 and Nedd4, to facilitate the budding process. These interactions serve as attractive targets for the development of broad-spectrum budding inhibitors. A major gap still exists in our understanding of the mechanism of filovirus budding due to the difficulty in detecting virus-host complexes and mapping their trafficking patterns in the natural environment of the cell. To address this gap, we used a bimolecular complementation (BiMC approach to detect, localize, and follow the trafficking patterns of eVP40-Tsg101 complexes in live mammalian cells. In addition, we used the BiMC approach along with a VLP budding assay to test small molecule inhibitors identified by in silico screening for their ability to block eVP40 PTAP-mediated interactions with Tsg101 and subsequent budding of eVP40 VLPs. We demonstrated the potential broad spectrum activity of a lead candidate inhibitor by demonstrating its ability to block PTAP-dependent binding of HIV-1 Gag to Tsg101 and subsequent egress of HIV-1 Gag VLPs.

  2. HVEM serial-section analysis of rabbit foliate taste buds: I. Type III cells and their synapses.

    Science.gov (United States)

    Royer, S M; Kinnamon, J C

    1991-04-01

    Serially sectioned rabbit foliate taste buds were examined with high voltage electron microscopy (HVEM) and computer-assisted, three-dimensional reconstruction. This report focuses on the ultrastructure of the type III cells and their synapses with sensory nerve fibers. Type III cells have previously been proposed to be the primary gustatory receptor cells in taste buds of rabbits and other mammals. Within rabbit foliate taste buds, type III cells constitute a well-defined, easily recognizable class and are the only taste bud cells observed to form synapses with intragemmal nerve fibers. Among 18 type III cells reconstructed from serial sections, 11 formed from 1 to 6 synapses each with nerve fibers; 7 reconstructed type III cells formed no synapses. Examples of both convergence and divergence of synaptic input from type III cells onto nerve fibers were observed. The sizes of the active zones of the synapses and numbers of vesicles associated with the presynaptic membrane specializations were highly variable. Dense-cored vesicles 80-140 nm in diameter were often found among the 40-60 nm clear vesicles clustered at presynaptic sites. At some synapses, these large dense-cored vesicles appeared to be the predominant vesicle type. This observation suggests that there may be functionally different types of synapses in taste buds, distinguished by the prevalence of either clear or dense-cored vesicles. Previous investigations have indicated that the dense-cored vesicles in type III cells may be storage sites for biogenic amines.

  3. Effect of Photoperiod Treatments on Dormancy Induction and Changes in Correlated Respiratory Rate of Nectarine Peach Bud

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The responses of dormancy induction to illumination and the characteristics of respiratory rate were studied with the nectarine peach bud in this article. The trial was conducted with nectarine (Prunus persica vat. nectariana cv. Shuguang) and involved three treatments: a short day treatment (8 h), a long day treatment (16 h), and the normal condition as the control. The dormancy status was determined with the growth of shoot and the sprouting ability, and the respiratory rate was mensurated with oxygen electrode. Short day treatment could induce the growth stopping of peach shoots ahead, promote the development of dormancy, and induce buds into dormancy with 21 d previous to control. Long day treatment postponed the growth stopping and the induction and development of dormancy. The respiratory rate decreased according to the development of dormancy induction. The minimum respiratory rate appeared about 7 days after the start of dormancy induction. Bud respiratory rate increased during this period and then declined and remained at low level during dormancy period. Long day reduced buds respiratory rate slightly. Short day could induce dormancy obviously, and long day postponed dormancy induction. The changes of respiratory rate were correlated with the development of dormancy induction, and the bud respiratory rate was also affected by photoperiod.

  4. Alteration of PHYA expression change circadian rhythms and timing of bud set in Populus.

    Science.gov (United States)

    Kozarewa, Iwanka; Ibáñez, Cristian; Johansson, Mikael; Ogren, Erling; Mozley, David; Nylander, Eva; Chono, Makiko; Moritz, Thomas; Eriksson, Maria E

    2010-05-01

    In many temperate woody species, dormancy is induced by short photoperiods. Earlier studies have shown that the photoreceptor phytochrome A (phyA) promotes growth. Specifically, Populus plants that over-express the oat PHYA gene (oatPHYAox) show daylength-independent growth and do not become dormant. However, we show that oatPHYAox plants could be induced to set bud and become cold hardy by exposure to a shorter, non-24 h diurnal cycle that significantly alters the relative position between endogenous rhythms and perceived light/dark cycles. Furthermore, we describe studies in which the expression of endogenous Populus tremula x P. tremuloides PHYTOCHROME A (PttPHYA) was reduced in Populus trees by antisense inhibition. The antisense plants showed altered photoperiodic requirements, resulting in earlier growth cessation and bud formation in response to daylength shortening, an effect that was explained by an altered innate period that leads to phase changes of clock-associated genes such as PttCO2. Moreover, gene expression studies following far-red light pulses show a phyA-mediated repression of PttLHY1 and an induction of PttFKF1 and PttFT. We conclude that the level of PttPHYA expression strongly influences seasonally regulated growth in Populus and is central to co-ordination between internal clock-regulated rhythms and external light/dark cycles through its dual effect on the pace of clock rhythms and in light signaling.

  5. New Flavonol Glucuronides from the Flower Buds of Syzygium aromaticum (Clove).

    Science.gov (United States)

    Ryu, Byeol; Kim, Hye Mi; Lee, Jin Su; Lee, Chan Kyu; Sezirahiga, Jurdas; Woo, Jeong-Hwa; Choi, Jung-Hye; Jang, Dae Sik

    2016-04-20

    Repeated chromatography of the EtOAc-soluble fraction from the 70% EtOH extract of the flower buds of Syzygium aromaticum (clove) led to the isolation and characterization of four new flavonol glucuronides, rhamnetin-3-O-β-d-glucuronide (1), rhamnazin-3-O-β-d-glucuronide (2), rhamnazin-3-O-β-d-glucuronide-6″-methyl ester (3), and rhamnocitrin-3-O-β-d-glucuronide-6″-methyl ester (4), together with 15 flavonoids (5-19) having previously known chemical structures. The structures of the new compounds 1-4 were determined by interpretation of spectroscopic data, particularly by 1D- and 2D-NMR studies. Six flavonoids (6, 7, 9, 14, 18, and 19) were isolated from the flower buds of S. aromaticum for the first time in this study. The flavonoids were examined for their cytotoxicity against human ovarian cancer cells (A2780) using MTT assays. Among the isolates, pachypodol (19) showed the most potent cytotoxicity on A2780 cells with an IC50 value of 8.02 μM. PMID:27045836

  6. Misexpression of Pknox2 in mouse limb bud mesenchyme perturbs zeugopod development and deltoid crest formation.

    Science.gov (United States)

    Zhou, Wenrong; Zhu, Huang; Zhao, Jianzhi; Li, Hanjun; Wan, Yong; Cao, Jingjing; Zhao, Haixia; Yu, Jian; Zhou, Rujiang; Yao, Yiyun; Zhang, Lingling; Wang, Lifang; He, Lin; Ma, Gang; Yao, Zhengju; Guo, Xizhi

    2013-01-01

    The TALE (Three Amino acid Loop Extension) family consisting of Meis, Pbx and Pknox proteins is a group of transcriptional co-factors with atypical homeodomains that play pivotal roles in limb development. Compared to the in-depth investigations of Meis and Pbx protein functions, the role of Pknox2 in limb development remains unclear. Here, we showed that Pknox2 was mainly expressed in the zeugopod domain of the murine limb at E10.5 and E11.5. Misexpression of Pknox2 in the limb bud mesenchyme of transgenic mice led to deformities in the zeugopod and forelimb stylopod deltoid crest, but left the autopod and other stylopod skeletons largely intact. These malformations in zeugopod skeletons were recapitulated in mice overexpressing Pknox2 in osteochondroprogenitor cells. Molecular and cellular analyses indicated that the misexpression of Pknox2 in limb bud mesenchyme perturbed the Hox10-11 gene expression profiles, decreased Col2 expression and Bmp/Smad signaling activity in the limb. These results indicated that Pknox2 misexpression affected mesenchymal condensation and early chondrogenic differentiation in the zeugopod skeletons of transgenic embryos, suggesting Pknox2 as a potential regulator of zeugopod and deltoid crest formation. PMID:23717575

  7. Misexpression of Pknox2 in mouse limb bud mesenchyme perturbs zeugopod development and deltoid crest formation.

    Directory of Open Access Journals (Sweden)

    Wenrong Zhou

    Full Text Available The TALE (Three Amino acid Loop Extension family consisting of Meis, Pbx and Pknox proteins is a group of transcriptional co-factors with atypical homeodomains that play pivotal roles in limb development. Compared to the in-depth investigations of Meis and Pbx protein functions, the role of Pknox2 in limb development remains unclear. Here, we showed that Pknox2 was mainly expressed in the zeugopod domain of the murine limb at E10.5 and E11.5. Misexpression of Pknox2 in the limb bud mesenchyme of transgenic mice led to deformities in the zeugopod and forelimb stylopod deltoid crest, but left the autopod and other stylopod skeletons largely intact. These malformations in zeugopod skeletons were recapitulated in mice overexpressing Pknox2 in osteochondroprogenitor cells. Molecular and cellular analyses indicated that the misexpression of Pknox2 in limb bud mesenchyme perturbed the Hox10-11 gene expression profiles, decreased Col2 expression and Bmp/Smad signaling activity in the limb. These results indicated that Pknox2 misexpression affected mesenchymal condensation and early chondrogenic differentiation in the zeugopod skeletons of transgenic embryos, suggesting Pknox2 as a potential regulator of zeugopod and deltoid crest formation.

  8. Interactions Between the Bud Rot Disease of Oil Palm and Rhynchophorus palmarum (Coleoptera: Curculionidae).

    Science.gov (United States)

    Plata-Rueda, Angelica; Martínez, Luis Carlos; Fernandes, Flávio Lemes; de Sousa Ramalho, Francisco; Zanuncio, José Cola; Serrão, José Eduardo

    2016-04-01

    Rhynchophorus palmarum (L.) causes great losses to the oil palm plantations, and therefore, the spatial and temporal distribution of this insect should be studied, to manage its populations. Insect sampling was done for 2 yr in an oil palm plantation from Colombia. In total, 60 pheromone traps were used in healthy palm trees and infected ones with the Bud Rot disease. On the other hand, developmental stages of this insect were quantified on healthy and diseased palms for two consecutive years. Number of adult R. palmarum per sampling was higher in the plantation with diseased palm trees, 3.85 and 74.7 insects per trap, than in those with healthy ones, 1.91 and 9.48 insects per trap, in the first and second years, respectively. After the integration of pheromone traps, there was a significant increase in the infestation level at all stages of development of the insect. For the first time, the presence of R. palmarum attracted to diseased palms is reported. The association between R. palmarum and the Bud Rot disease is a cause of death and great loss to the oil palm plantations. PMID:26791821

  9. Interactions Between the Bud Rot Disease of Oil Palm and Rhynchophorus palmarum (Coleoptera: Curculionidae).

    Science.gov (United States)

    Plata-Rueda, Angelica; Martínez, Luis Carlos; Fernandes, Flávio Lemes; de Sousa Ramalho, Francisco; Zanuncio, José Cola; Serrão, José Eduardo

    2016-04-01

    Rhynchophorus palmarum (L.) causes great losses to the oil palm plantations, and therefore, the spatial and temporal distribution of this insect should be studied, to manage its populations. Insect sampling was done for 2 yr in an oil palm plantation from Colombia. In total, 60 pheromone traps were used in healthy palm trees and infected ones with the Bud Rot disease. On the other hand, developmental stages of this insect were quantified on healthy and diseased palms for two consecutive years. Number of adult R. palmarum per sampling was higher in the plantation with diseased palm trees, 3.85 and 74.7 insects per trap, than in those with healthy ones, 1.91 and 9.48 insects per trap, in the first and second years, respectively. After the integration of pheromone traps, there was a significant increase in the infestation level at all stages of development of the insect. For the first time, the presence of R. palmarum attracted to diseased palms is reported. The association between R. palmarum and the Bud Rot disease is a cause of death and great loss to the oil palm plantations.

  10. Proteomic analysis of 'Zaosu' pear (Pyrus bretschneideri Rehd.) and its early-maturing bud sport.

    Science.gov (United States)

    Liu, Xueting; Zhai, Rui; Feng, Wenting; Zhang, Shiwei; Wang, Zhigang; Qiu, Zonghao; Zhang, Junke; Ma, Fengwang; Xu, Lingfei

    2014-07-01

    Maturation of fruits involves a series of physiological, biochemical, and organoleptic changes that eventually make fleshy fruits attractive, palatable, and nutritional. In order to understand the mature mechanism of the early-maturing bud sport of 'Zaosu' pear, we analyzed the differences of proteome expression between the both pears in different mature stages by the methods of a combination of two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis. Seventy-five differential expressed protein spots (psport, but only sixty-eight were demonstratively identified in the database of NCBI and uniprot. The majority of proteins were linked to metabolism, energy, stress response/defense and cell structure. Additionally, our data confirmed an increase of proteins related to cell-wall modification, oxidative stress and pentose phosphate metabolism and a decrease of proteins related to photosynthesis and glycolysis during the development process of both pears, but all these proteins increased or decreased faster in the early-maturing bud sport. This comparative analysis between both pears showed that these proteins were closely associated with maturation and could provide more detailed characteristics of the maturation process of both pears.

  11. Micropropagation of Codiaeum variegatum (L.) Blume and regeneration induction via adventitious buds and somatic embryogenesis.

    Science.gov (United States)

    Radice, Silvia

    2010-01-01

    Codiaeum variegatum (L) Blume cv. "Corazon de oro" and cv. "Norma" are successfully micropropagated when culture are initiated with explants taken from newly sprouted shoots. The establishment and multiplication steps are possible when 1 mg/L BA or 1 mg/L IAA and 3 mg/L 2iP are added to MS medium, according to the cultivar respectively selected.Adventive organogenesis and somatic embryogenesis are induced from leaf explants taken from in vitro buds of croton. On leaf-sectioned of "Corazon de oro" cultured in vitro, 1 mg/L BA stimulates continuous somatic embryos development and induces some shoots too. Replacing BA with 1 mg/L TDZ induces up to 100% bud regeneration in the same explants. On the other hand, leaf-sectioned of C. variegatum cv. Norma does not start somatic embryo differentiation if 1 mg/L TDZ is not added to the MS basal medium. Incipient callus is observed after 30 days of culture, and then, subculture to MS with 1 mg/L BA allows the same process to show on the "Corazon de oro" cultivar. Somatic embryos show growth arrest that is partially overcome by transfer to hormone-free basal medium with activated charcoal. Root induction is possible on basal medium plus 1 mg/L IBA. Plantlets in the greenhouse have variegated leaves true-to-type.

  12. Astral microtubule pivoting promotes their search for cortical anchor sites during mitosis in budding yeast.

    Directory of Open Access Journals (Sweden)

    Stephan Baumgärtner

    Full Text Available Positioning of the mitotic spindle is crucial for proper cell division. In the budding yeast Saccharomyces cerevisiae, two mechanisms contribute to spindle positioning. In the Kar9 pathway, astral microtubules emanating from the daughter-bound spindle pole body interact via the linker protein Kar9 with the myosin Myo2, which moves the microtubule along the actin cables towards the neck. In the dynein pathway, astral microtubules off-load dynein onto the cortical anchor protein Num1, which is followed by dynein pulling on the spindle. Yet, the mechanism by which microtubules target cortical anchor sites is unknown. Here we quantify the pivoting motion of astral microtubules around the spindle pole bodies, which occurs during spindle translocation towards the neck and through the neck. We show that this pivoting is largely driven by the Kar9 pathway. The microtubules emanating from the daughter-bound spindle pole body pivot faster than those at the mother-bound spindle pole body. The Kar9 pathway reduces the time needed for an astral microtubule inside the daughter cell to start pulling on the spindle. Thus, we propose a new role for microtubule pivoting: By pivoting around the spindle pole body, microtubules explore the space laterally, which helps them search for cortical anchor sites in the context of spindle positioning in budding yeast.

  13. Profiling DNA damage-induced phosphorylation in budding yeast reveals diverse signaling networks.

    Science.gov (United States)

    Zhou, Chunshui; Elia, Andrew E H; Naylor, Maria L; Dephoure, Noah; Ballif, Bryan A; Goel, Gautam; Xu, Qikai; Ng, Aylwin; Chou, Danny M; Xavier, Ramnik J; Gygi, Steven P; Elledge, Stephen J

    2016-06-28

    The DNA damage response (DDR) is regulated by a protein kinase signaling cascade that orchestrates DNA repair and other processes. Identifying the substrate effectors of these kinases is critical for understanding the underlying physiology and mechanism of the response. We have used quantitative mass spectrometry to profile DDR-dependent phosphorylation in budding yeast and genetically explored the dependency of these phosphorylation events on the DDR kinases MEC1, RAD53, CHK1, and DUN1. Based on these screens, a database containing many novel DDR-regulated phosphorylation events has been established. Phosphorylation of many of these proteins has been validated by quantitative peptide phospho-immunoprecipitation and examined for functional relevance to the DDR through large-scale analysis of sensitivity to DNA damage in yeast deletion strains. We reveal a link between DDR signaling and the metabolic pathways of inositol phosphate and phosphatidyl inositol synthesis, which are required for resistance to DNA damage. We also uncover links between the DDR and TOR signaling as well as translation regulation. Taken together, these data shed new light on the organization of DDR signaling in budding yeast. PMID:27298372

  14. Ingression Progression Complexes Control Extracellular Matrix Remodelling during Cytokinesis in Budding Yeast.

    Directory of Open Access Journals (Sweden)

    Magdalena Foltman

    2016-02-01

    Full Text Available Eukaryotic cells must coordinate contraction of the actomyosin ring at the division site together with ingression of the plasma membrane and remodelling of the extracellular matrix (ECM to support cytokinesis, but the underlying mechanisms are still poorly understood. In eukaryotes, glycosyltransferases that synthesise ECM polysaccharides are emerging as key factors during cytokinesis. The budding yeast chitin synthase Chs2 makes the primary septum, a special layer of the ECM, which is an essential process during cell division. Here we isolated a group of actomyosin ring components that form complexes together with Chs2 at the cleavage site at the end of the cell cycle, which we named 'ingression progression complexes' (IPCs. In addition to type II myosin, the IQGAP protein Iqg1 and Chs2, IPCs contain the F-BAR protein Hof1, and the cytokinesis regulators Inn1 and Cyk3. We describe the molecular mechanism by which chitin synthase is activated by direct association of the C2 domain of Inn1, and the transglutaminase-like domain of Cyk3, with the catalytic domain of Chs2. We used an experimental system to find a previously unanticipated role for the C-terminus of Inn1 in preventing the untimely activation of Chs2 at the cleavage site until Cyk3 releases the block on Chs2 activity during late mitosis. These findings support a model for the co-ordinated regulation of cell division in budding yeast, in which IPCs play a central role.

  15. Fission yeast hotspot sequence motifs are also active in budding yeast.

    Directory of Open Access Journals (Sweden)

    Walter W Steiner

    Full Text Available In most organisms, including humans, meiotic recombination occurs preferentially at a limited number of sites in the genome known as hotspots. There has been substantial progress recently in elucidating the factors determining the location of meiotic recombination hotspots, and it is becoming clear that simple sequence motifs play a significant role. In S. pombe, there are at least five unique sequence motifs that have been shown to produce hotspots of recombination, and it is likely that there are more. In S. cerevisiae, simple sequence motifs have also been shown to produce hotspots or show significant correlations with hotspots. Some of the hotspot motifs in both yeasts are known or suspected to bind transcription factors (TFs, which are required for the activity of those hotspots. Here we show that four of the five hotspot motifs identified in S. pombe also create hotspots in the distantly related budding yeast S. cerevisiae. For one of these hotspots, M26 (also called CRE, we identify TFs, Cst6 and Sko1, that activate and inhibit the hotspot, respectively. In addition, two of the hotspot motifs show significant correlations with naturally occurring hotspots. The conservation of these hotspots between the distantly related fission and budding yeasts suggests that these sequence motifs, and others yet to be discovered, may function widely as hotspots in many diverse organisms.

  16. Induction of callus from axillary buds of taro (Colocasia esculenta var. esculenta, Araceae) and subsequent plantlet regeneration.

    Science.gov (United States)

    Yam, T W; Young, J L; Fan, K P; Arditti, J

    1990-12-01

    Axillary buds of taro (Colocasia esculenta var. esculenta, Araceae) cultured on half strength Murashige-Skoog medium (HMS) containing taro extract (HMSTE) and 2, 4, 5-trichlorophenoxyacetic acid produce a compact, hard, slow growing callus which is not very active morphogenetically and produces only a few plantlets. When cultured on HMSTE plus 5 mg 1(-1) each of naphthaleneacetic acid and benzyl adenine (HMSNB) the buds produce a fast growing, friable and morphogenetically active callus. Meristematic regions form on the friable callus after 30 days on HMSNB. If transferred to HMSTE at this point the callus gives rise to plantlets. Addition of taro extract to the media is required for the culture of buds, induction of callus and plantlet regeneration.

  17. GDNF-independent ureteric budding: role of PI3K-independent activation of AKT and FOSB/JUN/AP-1 signaling

    Directory of Open Access Journals (Sweden)

    James B. Tee

    2013-07-01

    A significant fraction of mice deficient in either glial cell-derived neurotrophic factor (GDNF or its co-receptors (Gfrα1, Ret, undergoes ureteric bud (UB outgrowth leading to the formation of a rudimentary kidney. Previous studies using the isolated Wolffian duct (WD culture indicate that activation of fibroblast growth factor (FGF receptor signaling, together with suppression of BMP/Activin signaling, is critical for GDNF-independent WD budding (Maeshima et al., 2007. By expression analysis of embryonic kidney from Ret(−/− mice, we found the upregulation of several FGFs, including FGF7. To examine the intracellular pathways, we then analyzed GDNF-dependent and GDNF-independent budding in the isolated WD culture. In both conditions, Akt activation was found to be important; however, whereas this occurred through PI3-kinase in GDNF-dependent budding, in the case of GDNF-independent budding, Akt activation was apparently via a PI3-kinase independent mechanism. Jnk signaling and the AP-1 transcription factor complex were also implicated in GDNF-independent budding. FosB, a binding partner of c-Jun in the formation of AP-1, was the most highly upregulated gene in the ret knockout kidney (in which budding had still occurred, and we found that its siRNA-mediated knockdown in isolated WDs also blocked GDNF-independent budding. Taken together with the finding that inhibition of Jnk signaling does not block Akt activation/phosphorylation in GDNF-independent budding, the data support necessary roles for both FosB/Jun/AP-1 signaling and PI3-kinase-independent activation of Akt in GDNF-independent budding. A model is proposed for signaling events that involve Akt and JNK working to regulate GDNF-independent WD budding.

  18. Transcriptomic analysis of ‘Suli’ pear (Pyrus pyrifolia white pear group buds during the dormancy by RNA-Seq

    Directory of Open Access Journals (Sweden)

    Liu Guoqin

    2012-12-01

    Full Text Available Abstract Background Bud dormancy is a critical developmental process that allows perennial plants to survive unfavorable environmental conditions. Pear is one of the most important deciduous fruit trees in the world, but the mechanisms regulating bud dormancy in this species are unknown. Because genomic information for pear is currently unavailable, transcriptome and digital gene expression data for this species would be valuable resources to better understand the molecular and biological mechanisms regulating its bud dormancy. Results We performed de novo transcriptome assembly and digital gene expression (DGE profiling analyses of ‘Suli’ pear (Pyrus pyrifolia white pear group using the Illumina RNA-seq system. RNA-Seq generated approximately 100 M high-quality reads that were assembled into 69,393 unigenes (mean length = 853 bp, including 14,531 clusters and 34,194 singletons. A total of 51,448 (74.1% unigenes were annotated using public protein databases with a cut-off E-value above 10-5. We mainly compared gene expression levels at four time-points during bud dormancy. Between Nov. 15 and Dec. 15, Dec. 15 and Jan. 15, and Jan. 15 and Feb. 15, 1,978, 1,024, and 3,468 genes were differentially expressed, respectively. Hierarchical clustering analysis arranged 190 significantly differentially-expressed genes into seven groups. Seven genes were randomly selected to confirm their expression levels using quantitative real-time PCR. Conclusions The new transcriptomes offer comprehensive sequence and DGE profiling data for a dynamic view of transcriptomic variation during bud dormancy in pear. These data provided a basis for future studies of metabolism during bud dormancy in non-model but economically-important perennial species.

  19. Tissue culture of horse-radish (Cochlearia armoracia L. meristems: sterilization of buds and comparison of media

    Directory of Open Access Journals (Sweden)

    K. Górecka

    2015-06-01

    Full Text Available The attempt was made to cultivate horse-radish meristems taken from buds removed from roots. The lowest per cent of contamination was found after the buds had been soaiked in 80% ethanol for 6 minutes and then in 5%, 7.5% or 10% chloramine for 30, 30 or 15 minutes, respectively. Both agar media: Murashige-Skoog and Linsmaier-Skoog, were equally good, providing a moderate number of developing plants. The Linsmaier-Skoog medium was more satisfactory when solidified with agar; the results with liquid medium and filter-paper bridges were not as good.

  20. New biofunctional effects of the flower buds of Camellia sinensis and its bioactive acylated oleanane-type triterpene oligoglycosides.

    Science.gov (United States)

    Matsuda, Hisashi; Nakamura, Seikou; Morikawa, Toshio; Muraoka, Osamu; Yoshikawa, Masayuki

    2016-10-01

    We review the biofunctional effects of the flower buds of Camellia sinensis and C. sinensis var. assamica, such as antihyperlipidemic, antihyperglycemic, antiobesity, and gastroprotective effects in vivo, and antiallergic, pancreatic lipase inhibitory, and amyloid β (Aβ) aggregation inhibitory activities in vitro. Although the biofunctional effects of tea leaves have been extensively studied, less attention has been given to those of the flowers and seeds of the tea plant. Our studies focused on the saponin constituents of the extracts of the flower buds of C. sinensis cultivated in Japan and China, and C. sinensis var. assamica cultivated in India, and we review their beneficial biofunctions for health promotion.

  1. Lipopolysaccharide-induced inflammation attenuates taste progenitor cell proliferation and shortens the life span of taste bud cells

    Directory of Open Access Journals (Sweden)

    Brand Joseph

    2010-06-01

    Full Text Available Abstract Background The mammalian taste bud, a complex collection of taste sensory cells, supporting cells, and immature basal cells, is the structural unit for detecting taste stimuli in the oral cavity. Even though the cells of the taste bud undergo constant turnover, the structural homeostasis of the bud is maintained by balancing cell proliferation and cell death. Compared with nongustatory lingual epithelial cells, taste cells express higher levels of several inflammatory receptors and signalling proteins. Whether inflammation, an underlying condition in some diseases associated with taste disorders, interferes with taste cell renewal and turnover is unknown. Here we report the effects of lipopolysaccharide (LPS-induced inflammation on taste progenitor cell proliferation and taste bud cell turnover in mouse taste tissues. Results Intraperitoneal injection of LPS rapidly induced expression of several inflammatory cytokines, including tumor necrosis factor (TNF-α, interferon (IFN-γ, and interleukin (IL-6, in mouse circumvallate and foliate papillae. TNF-α and IFN-γ immunoreactivities were preferentially localized to subsets of cells in taste buds. LPS-induced inflammation significantly reduced the number of 5-bromo-2'-deoxyuridine (BrdU-labeled newborn taste bud cells 1-3 days after LPS injection, suggesting an inhibition of taste bud cell renewal. BrdU pulse-chase experiments showed that BrdU-labeled taste cells had a shorter average life span in LPS-treated mice than in controls. To investigate whether LPS inhibits taste cell renewal by suppressing taste progenitor cell proliferation, we studied the expression of Ki67, a cell proliferation marker. Quantitative real-time RT-PCR revealed that LPS markedly reduced Ki67 mRNA levels in circumvallate and foliate epithelia. Immunofluorescent staining using anti-Ki67 antibodies showed that LPS decreased the number of Ki67-positive cells in the basal regions surrounding circumvallate taste buds

  2. Consulting Centre for Substance Using Prevention in the Remand Prison in České Budějovice

    OpenAIRE

    ŠPÍRKOVÁ, Martina

    2012-01-01

    The thesis deals with the activities provided by the centre for drug prevention in the Remand Prison in České Budějovice. The theoretical part maps out the current drug issues within the society, the existing Czech prison system and the current drug issues in prisons observed. The practical part is focused on direct investigation of the activities provided by the centre for drug prevention in the Remand Prison in České Budějovice and clarification of what the prisoners have expected from this...

  3. Determination of the Effects of Endogenous Plant Hormones on alternate Bearing and Flower Bud Formation in Olives

    OpenAIRE

    ÜLGER, Salih; İbrahim BAKTIR; KAYNAK, Lami

    1998-01-01

    The effect of endogenous plant hormones on alternate-bearing, an important factor in olive growing, was investigated. Memecik and Tavşan Türeği olive cultivars were used. Leaf, bud, shoot tip and fruit samples were taken monthly during fruiting and non-fruiting periods for two years. Changes in ABA, GA3 and IAA were found and the effect of these hormones on the initiation of budding was determined. The results of the experiment showed that there were statistically important significances in ...

  4. Simulation of forest tree species' bud burst dates for different climate scenarios: chilling requirements and photo-period may limit bud burst advancement

    Science.gov (United States)

    Lange, Maximilian; Schaber, Jörg; Marx, Andreas; Jäckel, Greta; Badeck, Franz-Werner; Seppelt, Ralf; Doktor, Daniel

    2016-04-01

    This study investigates whether the assumed increase of winter and spring temperatures is depicted by phenological models in correspondingly earlier bud burst (BB) dates. Some studies assume that rising temperatures lead to an earlier BB, but even later BB has been detected. The phenological model PIM (promoter-inhibitor-model) fitted to the extensive phenological database of the German Weather Service was driven by several climate scenarios. This model accounts for the complicated mechanistic interactions between chilling requirements, temperature and photo-period. It predicts BB with a r 2 between 0.41 and 0.62 and a RMSE of around 1 week, depending on species. Parameter sensitivities depict species dependent interactions between growth and chilling requirements as well as photo-period. A mean trend to earlier BB was revealed for the period 2002- 2100, varying between -0.05 and -0.11 days per year, depending on species. These trends are lower than for the period 1951- 2009. Within climate scenario period, trends are decreasing for beech and chestnut, stagnating for birch and increasing for oak. Results suggest that not fulfilled chilling requirements accompanied by an increasing dependency on photo-period potentially limit future BB advancement. The combination of a powerful phenological model, a large scale phenological database and several climate scenarios, offers new insights into the mechanistic comprehension of spring phenology.

  5. Axillary budbreak in a cut rose crop as influenced by light intensity and red:far-red ratio at bud level

    NARCIS (Netherlands)

    Wubs-Timmermans, A.M.; Heuvelink, E.; Marcelis, L.F.M.; Buck-Sorlin, G.H.; Vos, J.

    2014-01-01

    When flower-bearing shoots in cut rose (Rosa ·hybrida) are harvested, a varying number of repressed axillary buds on the shoot remainder start to grow into new shoots (budbreak). Earlier experiments indicated that light reaching the bud affected the number of budbreaks. In all these studies, whole p

  6. LACK OF EXPRESSION OF EGF AND TGF-ALPHA IN THE FETAL MOUSE ALTERS FORMATION OF PROSTATIC EPITHELIAL BUDS AND INFLUENCES THE RESPONSE TO TCDD

    Science.gov (United States)

    Lack of Expression of EGF and TGF in the Fetal Mouse Alters Formation of Prostatic Epithelial Buds and Responsiveness to TCDD-Induced Impairment of Prostatic Bud Formation. Barbara D. Abbott, Tien-Min Lin, Nathan T. Rasmussen, Robert W. Moore,Ralph M. Albrecht, Judi...

  7. High floral bud abscission and lack of open flower abscission in Dendrobium cv. Miss Teen: rapid reduction of ethylene sensitivity in the abscission zone

    NARCIS (Netherlands)

    Bunya-atichart, K.; Ketsa, S.; Doorn, van W.G.

    2006-01-01

    We studied the abscission of floral buds and open flowers in cut Dendrobium inflorescences. Abscission of floral buds was high and sensitive to ethylene in all cultivars studied. Many open flowers abscised in most cultivars, but cv. Willie exhibited only small amount of floral fall and cv. Miss Teen

  8. Confinement to Organelle-Associated Inclusion Structures Mediates Asymmetric Inheritance of Aggregated Protein in Budding Yeast

    Directory of Open Access Journals (Sweden)

    Rachel Spokoini

    2012-10-01

    Full Text Available The division of the S. cerevisiae budding yeast, which produces one mother cell and one daughter cell, is asymmetric with respect to aging. Remarkably, the asymmetry of yeast aging coincides with asymmetric inheritance of damaged and aggregated proteins by the mother cell. Here, we show that misfolded proteins are retained in the mother cell by being sequestered in juxtanuclear quality control compartment (JUNQ and insoluble protein deposit (IPOD inclusions, which are attached to organelles. Upon exposure to stress, misfolded proteins accumulate in stress foci that must be disaggregated by Hsp104 in order to be degraded or processed to JUNQ and IPOD. Cells that fail to deliver aggregates to an inclusion pass on aggregates to subsequent generations.

  9. Anti-complement activity of tiliroside from the flower buds of Magnolia fargesii.

    Science.gov (United States)

    Jung, K Y; Oh, S R; Park, S H; Lee, I S; Ahn, K S; Lee, J J; Lee, H K

    1998-10-01

    As part of the search for anticomplementary active components from natural products, the anticomplementary properties of methanolic extracts from the flower buds of Magnoliafargesii have been investigated. Bioassay-guided chromatographic separation of the active constituents led to the isolation of compound 1, whose structure was identified by spectroscopic methods to be kaempferol 3-O-beta-D-(6"-O-coumaroyl)glucopyranoside (tiliroside). Tiliroside showed very potent anti-complement activity (IC50=5.4 x 10(-5) M) on the classical pathway of the complement system, even higher than rosmarinic acid, which is a well-known inhibitor against the complement system. On the other hand, the hydrolysates of tiliroside, kaempferol, astragalin and p-coumaric acid showed very weak activity on this system.

  10. Learning Shape and Texture Characteristics of CT Tree-in-Bud Opacities for CAD Systems

    CERN Document Server

    Bagci, Ulas; Caban, Jesus; Suffredini, Anthony F; Palmore, Tara N; Mollura, Daniel J

    2011-01-01

    Although radiologists can employ CAD systems to characterize malignancies, pulmonary fibrosis and other chronic diseases; the design of imaging techniques to quantify infectious diseases continue to lag behind. There exists a need to create more CAD systems capable of detecting and quantifying characteristic patterns often seen in respiratory tract infections such as influenza, bacterial pneumonia, or tuborculosis. One of such patterns is Tree-in-bud (TIB) which presents \\textit{thickened} bronchial structures surrounding by clusters of \\textit{micro-nodules}. Automatic detection of TIB patterns is a challenging task because of their weak boundary, noisy appearance, and small lesion size. In this paper, we present two novel methods for automatically detecting TIB patterns: (1) a fast localization of candidate patterns using information from local scale of the images, and (2) a M\\"{o}bius invariant feature extraction method based on learned local shape and texture properties. A comparative evaluation of the pr...

  11. Identification of cellular factors required for the budding of koala retrovirus.

    Science.gov (United States)

    Shimode, Sayumi; Nakaoka, Rie; Hoshino, Shigeki; Abe, Masumi; Shogen, Hiroko; Yasuda, Jiro; Miyazawa, Takayuki

    2013-07-01

    Koala retrovirus (KoRV) is a unique gammaretrovirus that is currently endogenizing into its host and considered to be associated with leukemia, lymphoma and immunosuppression in koalas (Phascolactos cinereus). In this study, it was demonstrated that WWP2 or WWP2-like E3 ubiquitin ligases possessing the WW domain closely related to WWP2 and Vps4A/B are involved in KoRV budding. These data suggest that KoRV Gag recruits the cellular endosomal sorting complex required for transport machinery through interaction of the PPPY L-domain with the WW domain(s) of WWP2 and that progeny virions are released from cells by utilizing the multivesicular body sorting pathway.

  12. Development of automatic image analysis algorithms for protein localization studies in budding yeast

    Science.gov (United States)

    Logg, Katarina; Kvarnström, Mats; Diez, Alfredo; Bodvard, Kristofer; Käll, Mikael

    2007-02-01

    Microscopy of fluorescently labeled proteins has become a standard technique for live cell imaging. However, it is still a challenge to systematically extract quantitative data from large sets of images in an unbiased fashion, which is particularly important in high-throughput or time-lapse studies. Here we describe the development of a software package aimed at automatic quantification of abundance and spatio-temporal dynamics of fluorescently tagged proteins in vivo in the budding yeast Saccharomyces cerevisiae, one of the most important model organisms in proteomics. The image analysis methodology is based on first identifying cell contours from bright field images, and then use this information to measure and statistically analyse protein abundance in specific cellular domains from the corresponding fluorescence images. The applicability of the procedure is exemplified for two nuclear localized GFP-tagged proteins, Mcm4p and Nrm1p.

  13. From assembly to virus particle budding: pertinence of the detergent resistant membranes.

    Science.gov (United States)

    Gosselin-Grenet, Anne-Sophie; Mottet-Osman, Geneviève; Roux, Laurent

    2006-01-20

    Detergent resistant membranes (DRMs) are the site of assembly for a variety of viruses. Here, we make use of Sendai virus mutant proteins that are not packaged into virus particles to determine the involvement of this assembly for the virus particle production. We found that, in the context of an infection, (1) all the Sendai virus proteins associated in part with DRMs, (2) mutant HN and M proteins not packaged into virus particles were similarly part of this association, (3) after M protein suppression resulting in a significant reduction of virus production, the floatation profile of the other viral proteins was not altered and finally (4) cellular cholesterol depletion did not decrease the virus particle production, although it somehow reduced their virus infectivity. These results led us to conclude that the assembly complex found in DRM fractions does not constitute a direct precursor of virus particle budding.

  14. Sequential Feedback Induction Stabilizes the Phosphate Starvation Response in Budding Yeast

    Directory of Open Access Journals (Sweden)

    Noam Vardi

    2014-11-01

    Full Text Available Depletion of essential nutrients triggers regulatory programs that prolong cell growth and survival. Starvation-induced processes increase nutrient transport, mobilize nutrient storage, and recycle nutrients between cellular components. This leads to an effective increase in intracellular nutrients, which may act as a negative feedback that downregulates the starvation program. To examine how cells overcome this potential instability, we followed the transcription response of budding yeast transferred to medium lacking phosphate. Genes were induced in two temporal waves. The first wave was stably maintained and persisted even upon phosphate replenishment, indicating a positive feedback loop. This commitment was abolished after 2 hr with the induction of the second expression wave, coinciding with the reduction in cell growth rate. We show that the overall temporal stability of the expression response depends on the sequential pattern of gene induction. Our results emphasize the key role of gene expression dynamics in optimizing cellular adaptation.

  15. A microfluidic system for studying ageing and dynamic single-cell responses in budding yeast.

    Directory of Open Access Journals (Sweden)

    Matthew M Crane

    Full Text Available Recognition of the importance of cell-to-cell variability in cellular decision-making and a growing interest in stochastic modeling of cellular processes has led to an increased demand for high density, reproducible, single-cell measurements in time-varying surroundings. We present ALCATRAS (A Long-term Culturing And TRApping System, a microfluidic device that can quantitatively monitor up to 1000 cells of budding yeast in a well-defined and controlled environment. Daughter cells are removed by fluid flow to avoid crowding allowing experiments to run for over 60 hours, and the extracellular media may be changed repeatedly and in seconds. We illustrate use of the device by measuring ageing through replicative life span curves, following the dynamics of the cell cycle, and examining history-dependent behaviour in the general stress response.

  16. ultrastructural study of limb bud development in green turtles chelonia mydas

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    morphological changes during the embryonic development of limbs of the green turtle,chelonia mydas,were studied during the entire period of incubation,using transmission and scanning electron microscopy (tem and sem).limb buds were first observed at stage 2.at that stage,the tip was covered with an apical ectodermal ridge (aer) which began to regress at stage 6.associated with aer was the presence of the mesenchymal cells which,consequently,differentiated into muscles,cartilage and bones.the gross features of the skeletal development appeared as a condensation of the cartilaginous structures in the proximal distal region of the limbs.the primordial digits were gradually enclosed by hard keratinized webbed skin.the increase in rate of ossification and skin pigmentation was correlated with the growth of the limbs.the development of the limbs was closely related to the transitional appearance of mucus secretion from the epidermis.

  17. Bionomy of spruce bud scale, Physokermes piceae (schrank (Hemiptera: Coccidae in the Belgrade area, Serbia

    Directory of Open Access Journals (Sweden)

    Graora Draga

    2012-01-01

    Full Text Available Spruce bud scale, Physokermes piceae, develops on the genus Picea. Large colonies of this species are constantly present on Picea abies in green areas in the Belgrade territory, causing the drying of needles, branches and whole plants. Therefore, Ph. piceae is a significant spruce pest. Spruce scales attract many entomophagous insects able to reduce pest population. Parasitoid wasps Coccophagus lycimnia (Walk (Aphelinidae and Microterys lunatus (Dalm. (Encyrtidae were reared. Predators Exochomus quadripustulatus L., Scymnus abietis Paykull (Coccinellidae and Anthribus nebulosus Forster (Anthribidae were determined. Both species of ladybird were confirmed as predators of Ph. piceae for the first time in Serbia, while S. abietis is a new species for the fauna of Serbia. The most effective natural enemy of Ph. piceae was A. nebulosus, reducing populations by 68-80%.

  18. AGROBACTERIUM MEDIATED TRANSFORMATION OF PIGEONPEA (CAJANUS CAJAN L MILLLSP VAR LRG-41 FROM AXILLARY BUD

    Directory of Open Access Journals (Sweden)

    T. Raghavendra

    2014-02-01

    Full Text Available A reliable method of plant regeneration has been achieved from Axillary buds. Shoots appeared from explants when cultured on Murashige and skoog (MS medium supplemented with BAP (Benzyl amino purine, Napthalene acetic acid (NAA and Kinetin at various combinations. Elongated shoots were rooted with 70.6% rooting frequency in MS medium with indole buteric acid (IBA at 1.0mg/l. The rooted plantlets were established well in soilrite mixture medium with 91% success and days taken for acclimatization were 12.8 days. This protocol was used in Agrobacterium mediated transformation. The transformation was carried out using the Agrobacterium strain LBA4404 containing the binary vector pCAMBIA2301 harboring npt II as selectable marker and GUS as reporter gene.

  19. AGROBACTERIUM MEDIATED TRANSFORMATION OF PIGEONPEA (CAJANUS CAJAN L MILLLSP VAR LRG-41 FROM AXILLARY BUD

    Directory of Open Access Journals (Sweden)

    T. Raghavendra

    2014-03-01

    Full Text Available A reliable method of plant regeneration has been achieved from Axillary buds. Shoots appeared from explants when cultured on Murashige and skoog (MS medium supplemented with BAP (Benzyl amino purine, Napthalene acetic acid (NAA and Kinetin at various combinations. Elongated shoots were rooted with 70.6% rooting frequency in MS medium with indole buteric acid (IBA at 1.0mg/l. The rooted plantlets were established well in soilrite mixture medium with 91% success and days taken for acclimatization were 12.8 days. This protocol was used in Agrobacterium mediated transformation. The transformation was carried out using the Agrobacterium strain LBA4404 containing the binary vector pCAMBIA2301 harboring npt II as selectable marker and GUS as reporter gene.

  20. Use of active extracts of poplar buds against Penicillium italicum and possible modes of action.

    Science.gov (United States)

    Yang, Shuzhen; Liu, Limei; Li, Dongmei; Xia, Huan; Su, Xiaojun; Peng, Litao; Pan, Siyi

    2016-04-01

    Antifungal components, from poplar buds active fraction (PBAF) against Penicillium italicum, the causal agent of blue mold in citrus fruits, were identified and possible action modes were investigated. Pinocembrin, chrysin and galangin were determined as active components in PBAF, using HPLC and HPLC-MS analysis. The antifungal activity is stable at temperatures ranging from 4 °C to 100 °C and pH levels ranging from 4 to 8. In the presence of PBAF, the hyphae become shriveled, wrinkled and the cell membrane became seriously disrupted. Further investigation on cell permeability, nucleic acid content and alkaline phosphatase suggest that the cell membrane might be the target. Mycelial oxygen consumption and the respiration-related enzymatic activity of succinate dehydrogenase, malate dehydrogenase and ATPase were all inhibited by PBAF. We propose that PBAF is a potentially useful alternative for blue mold control and may act against P. italicum by interfering with respiration and disrupting the cell membrane. PMID:26593534

  1. Identification of the Flavone in the Flower Bud of Panax Quinquefolium and Its Content Determination in the Different Parts

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@Introduction   Panax quinquefolium L. Belongs to the Panax genus of the Araliaceae. It originates in America and Canada. Its roots, a famous and expensive traditional Chinese medicinal material, have been studied continually since its successful cultivation in China in the 1970s. In recent years the development and utilization researches of its aerial part, which was abandoned in the past, have been conducted. The constituent studies on the leaves, stems and fruits have been reported. But no information about the flower buds has been found in literatures. The authors have reported a study about the chemical constituents of the flower buds of the P. Quinquefolium cultivated in China. In the present work two flavonoids(namely kaempferol and panasenoside) in the flower buds of the P. Quinquefulium cultivated in China were extracted, isolated, and identified. Since the medicinal materials were processed with an unusual treatment, the content determination of the monomeric and total flavonoid in the different parts were performed correctly with the methods of dual-wavelength TLC-scan, and UV-absorption, and it was discovered that there were flavonoids in the roots and fruits of the P. Quinquefolium. The scientific basis was provided for the utilization of the flower buds and other parts resources of the P.

  2. Extended Low Temperature Impacts Dormancy Status, Flowering Competence, and Transcript Profiles in Crown Buds of Leafy Spurge

    Science.gov (United States)

    Leafy spurge (Euphorbia esula) is an herbaceous perennial weed that reproduces vegetatively from an abundance of underground adventitious buds. In this study we report the effects of different growth conditions on vegetative reproduction and flowering competence, and determine molecular mechanisms a...

  3. Update History of This Database - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available [ Credits ] BLAST Search Image Search Home About Archive Update History Contact us ...Budding yeast cDNA sequencing project Update History of This Database Date Update contents 2010/03/29 Buddin...tio About This Database Database Description Download License Update History of This Database Site Policy | Contact Us Update

  4. Dormancy of 'Imperial Gala' apple and 'Hosui' pear tree buds in a region of low chill occurrence

    Directory of Open Access Journals (Sweden)

    Ruy Inácio Neiva de Carvalho

    2014-08-01

    Full Text Available The objective of this work was to evaluate the dormancy dynamic of Imperial Gala apple tree buds and Hosui pear tree buds in a region of low chill occurrence. Experiments were conducted between April and August in 2007 and 2008. Branches were collected every two weeks from an orchard at Porto Amazonas (Paraná State, Brazil. On the last sampling day, an additional set of branches was collected and refrigerated between 4°C and 7°C for 1,440 hours. Dormancy was evaluated using a biological test of single node cuttings isolated in growth chambers (GC at 25°C with 16 hours of light exposure. The number of chill hours (CH and chill units (CU for the region were recorded. The two species were evaluated in separate experiments. We used 11 completely randomized treatments with four replicas for each species. The peak of endodormancy for the Imperial Gala apple tree buds occurred in early June 2007 and from middle June to early July in 2008. The endodormancy of the Hosui pear tree buds oscillated between April and August in 2007 and peaked between June and early July in 2008.

  5. Effects of Allspice, Cinnamon, and Clove Bud Essential Oils in Edible Apple Films on Physical Properties and Antimicrobial Activities

    Science.gov (United States)

    The results of the present study show that allspice, cinnamon and clove bud essential oils can be used to prepare apple-based antimicrobial edible films with good physical properties for food applications by both direct contact and indirectly by vapors emanating from the films. Application of the a...

  6. Measurement of the volume growth rate of single budding yeast with the MOSFET-based microfluidic Coulter counter.

    Science.gov (United States)

    Sun, Jiashu; Stowers, Chris C; Boczko, Erik M; Li, Deyu

    2010-11-01

    We report on measurements of the volume growth rate of ten individual budding yeast cells using a recently developed MOSFET-based microfluidic Coulter counter. The MOSFET-based microfluidic Coulter counter is very sensitive, provides signals that are immune from the baseline drift, and can work with cell culture media of complex composition. These desirable features allow us to directly measure the volume growth rate of single cells of Saccharomyces cerevisiae LYH3865 strain budding yeast in YNB culture media over a whole cell cycle. Results indicate that all budding yeast follow a sigmoid volume growth profile with reduced growth rates at the initial stage before the bud emerges and the final stage after the daughter gets mature. Analysis of the data indicates that even though all piecewise linear, Gomperitz, and Hill's function models can fit the global growth profile equally well, the data strongly support local exponential growth phenomenon. Accurate volume growth measurements are important for applications in systems biology where quantitative parameters are required for modeling and simulation. PMID:20717618

  7. Changes of the Unique Odontogenic Properties of Rat Apical Bud Cells under the Developing Apical Complex Microenvironment

    Institute of Scientific and Technical Information of China (English)

    Jun Fang; Liang Tang; Xiao-hui Liu; Ling-ying Wen; Yan Jin

    2009-01-01

    Aim To characterize the odontogenic capability of apical bud and phenotypical change of apical bud cells (ABCs) in different microenvironment. Methodology Incisor apical bud tissues from neonatal SD rat were dissected and transplanted into the renal capsules to determine their odontogenic capability. Meanwhile ABCs were cultured and purified by repeated differential trypsinization. Then ABCs were cultured with conditioned medium from developing apical complex cells (DAC-CM). Immunocytochemistry, reverse transcriptase polymerase chain reaction (RT-PCR) and scanning electron microscope (SEM) were performed to compare the biological change of ABC treated with or without DAC-CM. Results First we confirmed the ability of apical bud to form crown-like structure ectopically. Equally important, by using the developing apical complex (DAC) conditioned medium, we found the microenvironment created by root could abrogate the "crown" features of ABCs and promote their proliferation and differentiation. Conclusion ABCs possess odontogenic capability to form crown-like tissues and this property can be affected by root-produced microenvironment.

  8. Evidence for a role of glutamate as an efferent transmitter in taste buds

    Directory of Open Access Journals (Sweden)

    Anderson Catherine B

    2010-06-01

    Full Text Available Abstract Background Glutamate has been proposed as a transmitter in the peripheral taste system in addition to its well-documented role as an umami taste stimulus. Evidence for a role as a transmitter includes the presence of ionotropic glutamate receptors in nerve fibers and taste cells, as well as the expression of the glutamate transporter GLAST in Type I taste cells. However, the source and targets of glutamate in lingual tissue are unclear. In the present study, we used molecular, physiological and immunohistochemical methods to investigate the origin of glutamate as well as the targeted receptors in taste buds. Results Using molecular and immunohistochemical techniques, we show that the vesicular transporters for glutamate, VGLUT 1 and 2, but not VGLUT3, are expressed in the nerve fibers surrounding taste buds but likely not in taste cells themselves. Further, we show that P2X2, a specific marker for gustatory but not trigeminal fibers, co-localizes with VGLUT2, suggesting the VGLUT-expressing nerve fibers are of gustatory origin. Calcium imaging indicates that GAD67-GFP Type III taste cells, but not T1R3-GFP Type II cells, respond to glutamate at concentrations expected for a glutamate transmitter, and further, that these responses are partially blocked by NBQX, a specific AMPA/Kainate receptor antagonist. RT-PCR and immunohistochemistry confirm the presence of the Kainate receptor GluR7 in Type III taste cells, suggesting it may be a target of glutamate released from gustatory nerve fibers. Conclusions Taken together, the results suggest that glutamate may be released from gustatory nerve fibers using a vesicular mechanism to modulate Type III taste cells via GluR7.

  9. BuD, a helix–loop–helix DNA-binding domain for genome modification

    Energy Technology Data Exchange (ETDEWEB)

    Stella, Stefano [Spanish National Cancer Research Centre (CNIO), Calle de Melchor Fernández Almagro 3, 28029 Madrid (Spain); University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen (Denmark); Molina, Rafael; López-Méndez, Blanca [Spanish National Cancer Research Centre (CNIO), Calle de Melchor Fernández Almagro 3, 28029 Madrid (Spain); Juillerat, Alexandre; Bertonati, Claudia; Daboussi, Fayza [Cellectis, 8 Rue de la Croix Jarry, 75013 Paris (France); Campos-Olivas, Ramon [Spanish National Cancer Research Centre (CNIO), Calle de Melchor Fernández Almagro 3, 28029 Madrid (Spain); Duchateau, Phillippe [Cellectis, 8 Rue de la Croix Jarry, 75013 Paris (France); Montoya, Guillermo, E-mail: guillermo.montoya@cpr.ku.dk [Spanish National Cancer Research Centre (CNIO), Calle de Melchor Fernández Almagro 3, 28029 Madrid (Spain); University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen (Denmark)

    2014-07-01

    Crystal structures of BurrH and the BurrH–DNA complex are reported. DNA editing offers new possibilities in synthetic biology and biomedicine for modulation or modification of cellular functions to organisms. However, inaccuracy in this process may lead to genome damage. To address this important problem, a strategy allowing specific gene modification has been achieved through the addition, removal or exchange of DNA sequences using customized proteins and the endogenous DNA-repair machinery. Therefore, the engineering of specific protein–DNA interactions in protein scaffolds is key to providing ‘toolkits’ for precise genome modification or regulation of gene expression. In a search for putative DNA-binding domains, BurrH, a protein that recognizes a 19 bp DNA target, was identified. Here, its apo and DNA-bound crystal structures are reported, revealing a central region containing 19 repeats of a helix–loop–helix modular domain (BurrH domain; BuD), which identifies the DNA target by a single residue-to-nucleotide code, thus facilitating its redesign for gene targeting. New DNA-binding specificities have been engineered in this template, showing that BuD-derived nucleases (BuDNs) induce high levels of gene targeting in a locus of the human haemoglobin β (HBB) gene close to mutations responsible for sickle-cell anaemia. Hence, the unique combination of high efficiency and specificity of the BuD arrays can push forward diverse genome-modification approaches for cell or organism redesign, opening new avenues for gene editing.

  10. Tumor budding as a potential histopathological biomarker in colorectal cancer: Hype or hope?

    Institute of Scientific and Technical Information of China (English)

    Fabio Grizzi; Giuseppe Celesti; Gianluca Basso; Luigi Laghi

    2012-01-01

    Colorectal cancer (CRC),the third most commonly diagnosed type of cancer in men and women worldwide is recognized as a complex multi-pathway disease,an observation sustained by the fact that histologically identical tumors may have different outcome,including various response to therapy.Therefore,particularly in early and intermediate stage (stages Ⅱ and Ⅲ,respectively) CRC,there is a compelling need for biomarkers helpful of selecting patients with aggressive disease that might benefit from adjuvant and targeted therapy.Histopathological examination shows that likely other solid tumors the development and progression of human CRC is not only determined by genetically abnormal cells,but also by intricate interactions between malignant cells and the surrounding microenvironment.This has led to reconsider the features of tumor microenvironment as potential predictive and prognostic biomarkers.Among the histopathological biomarkers,tumor budding (i.e.,the presence of individual cells and small clusters of tumor cells at the tumor invasive front)has received much recent attention,particularly in the setting of CRC.Although its acceptance as a reportable factor has been held back by a lack of uniformity with respect to qualitative and quantitative aspects,tumor budding is now considered as an independent adverse prognostic factor in CRC that may allow for stratification of patients into risk categories more meaningful than those defined by tumor-node-metastasis staging alone,and also potentially guide treatment decisions,especially in T2-T3 NO (stage Ⅱ) CRCs.

  11. BuD, a helix–loop–helix DNA-binding domain for genome modification

    International Nuclear Information System (INIS)

    Crystal structures of BurrH and the BurrH–DNA complex are reported. DNA editing offers new possibilities in synthetic biology and biomedicine for modulation or modification of cellular functions to organisms. However, inaccuracy in this process may lead to genome damage. To address this important problem, a strategy allowing specific gene modification has been achieved through the addition, removal or exchange of DNA sequences using customized proteins and the endogenous DNA-repair machinery. Therefore, the engineering of specific protein–DNA interactions in protein scaffolds is key to providing ‘toolkits’ for precise genome modification or regulation of gene expression. In a search for putative DNA-binding domains, BurrH, a protein that recognizes a 19 bp DNA target, was identified. Here, its apo and DNA-bound crystal structures are reported, revealing a central region containing 19 repeats of a helix–loop–helix modular domain (BurrH domain; BuD), which identifies the DNA target by a single residue-to-nucleotide code, thus facilitating its redesign for gene targeting. New DNA-binding specificities have been engineered in this template, showing that BuD-derived nucleases (BuDNs) induce high levels of gene targeting in a locus of the human haemoglobin β (HBB) gene close to mutations responsible for sickle-cell anaemia. Hence, the unique combination of high efficiency and specificity of the BuD arrays can push forward diverse genome-modification approaches for cell or organism redesign, opening new avenues for gene editing

  12. A hybrid agent-based model of the developing mammary terminal end bud.

    Science.gov (United States)

    Butner, Joseph D; Chuang, Yao-Li; Simbawa, Eman; Al-Fhaid, A S; Mahmoud, S R; Cristini, Vittorio; Wang, Zhihui

    2016-10-21

    Mammary gland ductal elongation is spearheaded by terminal end buds (TEBs), where populations of highly proliferative cells are maintained throughout post-pubertal organogenesis in virgin mice until the mammary fat pad is filled by a mature ductal tree. We have developed a hybrid multiscale agent-based model to study how cellular differentiation pathways, cellular proliferation capacity, and endocrine and paracrine signaling play a role during development of the mammary gland. A simplified cellular phenotypic hierarchy that includes stem, progenitor, and fully differentiated cells within the TEB was implemented. Model analysis finds that mammary gland development was highly sensitive to proliferation events within the TEB, with progenitors likely undergoing 2-3 proliferation cycles before transitioning to a non-proliferative phenotype, and this result is in agreement with our previous experimental work. Endocrine and paracrine signaling were found to provide reliable ductal elongation rate regulation, while variations in the probability a new daughter cell will be of a proliferative phenotype were seen to have minimal effects on ductal elongation rates. Moreover, the distribution of cellular phenotypes within the TEB was highly heterogeneous, demonstrating significant allowable plasticity in possible phenotypic distributions while maintaining biologically relevant growth behavior. Finally, simulation results indicate ductal elongation rates due to cellular proliferation within the TEB may have a greater sensitivity to upstream endocrine signaling than endothelial to stromal paracrine signaling within the TEB. This model provides a useful tool to gain quantitative insights into cellular population dynamics and the effects of endocrine and paracrine signaling within the pubertal terminal end bud. PMID:27475843

  13. Differential expression of a BMP4 reporter allele in anterior fungiform versus posterior circumvallate taste buds of mice

    Directory of Open Access Journals (Sweden)

    Barlow Linda A

    2010-10-01

    Full Text Available Abstract Background Bone Morphogenetic Protein 4 (BMP4 is a diffusible factor which regulates embryonic taste organ development. However, the role of BMP4 in taste buds of adult mice is unknown. We utilized transgenic mice with LacZ under the control of the BMP4 promoter to reveal the expression of BMP4 in the tongues of adult mice. Further we evaluate the pattern of BMP4 expression with that of markers of specific taste bud cell types and cell proliferation to define and compare the cell populations expressing BMP4 in anterior (fungiform papillae and posterior (circumvallate papilla tongue. Results BMP4 is expressed in adult fungiform and circumvallate papillae, i.e., lingual structures composed of non-taste epithelium and taste buds. Unexpectedly, we find both differences and similarities with respect to expression of BMP4-driven ß-galactosidase. In circumvallate papillae, many fusiform cells within taste buds are BMP4-ß-gal positive. Further, a low percentage of BMP4-expressing cells within circumvallate taste buds is immunopositive for markers of each of the three differentiated taste cell types (I, II and III. BMP4-positive intragemmal cells also expressed a putative marker of immature taste cells, Sox2, and consistent with this finding, intragemmal cells expressed BMP4-ß-gal within 24 hours after their final mitosis, as determined by BrdU birthdating. By contrast, in fungiform papillae, BMP4-ß-gal positive cells are never encountered within taste buds. However, in both circumvallate and fungiform papillae, BMP4-ß-gal expressing cells are located in the perigemmal region, comprising basal and edge epithelial cells adjacent to taste buds proper. This region houses the proliferative cell population that gives rise to adult taste cells. However, perigemmal BMP4-ß-gal cells appear mitotically silent in both fungiform and circumvallate taste papillae, as we do not find evidence of their active proliferation using cell cycle immunomarkers

  14. Synthetic retinoids, retinobenzoic acids, Am80, Am580 and Ch55 regulate morphogenesis in chick limb bud.

    Science.gov (United States)

    Tamura, K; Kagechika, H; Hashimoto, Y; Shudo, K; Ohsugi, K; Ide, H

    1990-10-01

    The retinobenzoic acids Am80, Am580 and Ch55 are synthetic stable analogs of retinoic acid (RA), and show very strong differentiation-inducing activity in human myelogeneous leukemia cell line HL-60. To examine the effects of these synthetic retinoids on limb pattern formation, AG1-X2 beads containing these retinoids were applied to the anterior margin of stage 19-20 chick wing buds. By implanting the beads with 1 microgram/ml retinoids, normal wings were formed and extra digits 2 or 32 were rarely formed. As the retinoid concentrations increased from 10 micrograms/ml to 100 micrograms/ml duplicated limbs 3234, 43234, 432234, 4334 were progressively produced. At higher concentrations, 1 mg/ml, the wings often truncated, although duplication occurred in some embryos. These synthetic analogs seem to have the same degree of morphogenetic potential as RA, since the activity index of these retinoids was similar to that of RA. Since these synthetic retinoids hardly bind to CRABP (cellular retinoic acid-binding protein), it may be possible that the retinoids and RA may affect limb-pattern formation without the interaction with CRABP. It is known that limb buds cannot develop distal structures when the posterior region including all ZPA (zone of polarizing activity) is removed. When beads containing the above mentioned retinoids were implanted to the anterior margin of wing buds from which the posterior one third region including all ZPA had been removed, distal growth of the wing buds and the formation of digit elements were observed. Some of the wing buds produced a completely reverse digit pattern 432. From these results, we discussed the roles of RA in limb development and pattern formation.

  15. A secreted BMP antagonist, Cer1, fine tunes the spatial organization of the ureteric bud tree during mouse kidney development.

    Directory of Open Access Journals (Sweden)

    Lijun Chi

    Full Text Available The epithelial ureteric bud is critical for mammalian kidney development as it generates the ureter and the collecting duct system that induces nephrogenesis in dicrete locations in the kidney mesenchyme during its emergence. We show that a secreted Bmp antagonist Cerberus homologue (Cer1 fine tunes the organization of the ureteric tree during organogenesis in the mouse embryo. Both enhanced ureteric expression of Cer1 and Cer1 knock out enlarge kidney size, and these changes are associated with an altered three-dimensional structure of the ureteric tree as revealed by optical projection tomography. Enhanced Cer1 expression changes the ureteric bud branching programme so that more trifid and lateral branches rather than bifid ones develop, as seen in time-lapse organ culture. These changes may be the reasons for the modified spatial arrangement of the ureteric tree in the kidneys of Cer1+ embryos. Cer1 gain of function is associated with moderately elevated expression of Gdnf and Wnt11, which is also induced in the case of Cer1 deficiency, where Bmp4 expression is reduced, indicating the dependence of Bmp expression on Cer1. Cer1 binds at least Bmp2/4 and antagonizes Bmp signalling in cell culture. In line with this, supplementation of Bmp4 restored the ureteric bud tip number, which was reduced by Cer1+ to bring it closer to the normal, consistent with models suggesting that Bmp signalling inhibits ureteric bud development. Genetic reduction of Wnt11 inhibited the Cer1-stimulated kidney development, but Cer1 did not influence Wnt11 signalling in cell culture, although it did inhibit the Wnt3a-induced canonical Top Flash reporter to some extent. We conclude that Cer1 fine tunes the spatial organization of the ureteric tree by coordinating the activities of the growth-promoting ureteric bud signals Gndf and Wnt11 via Bmp-mediated antagonism and to some degree via the canonical Wnt signalling involved in branching.

  16. Fruit load induces changes in global gene expression and in abscisic acid (ABA) and indole acetic acid (IAA) homeostasis in citrus buds.

    Science.gov (United States)

    Shalom, Liron; Samuels, Sivan; Zur, Naftali; Shlizerman, Lyudmila; Doron-Faigenboim, Adi; Blumwald, Eduardo; Sadka, Avi

    2014-07-01

    Many fruit trees undergo cycles of heavy fruit load (ON-Crop) in one year, followed by low fruit load (OFF-Crop) the following year, a phenomenon known as alternate bearing (AB). The mechanism by which fruit load affects flowering induction during the following year (return bloom) is still unclear. Although not proven, it is commonly accepted that the fruit or an organ which senses fruit presence generates an inhibitory signal that moves into the bud and inhibits apical meristem transition. Indeed, fruit removal from ON-Crop trees (de-fruiting) induces return bloom. Identification of regulatory or metabolic processes modified in the bud in association with altered fruit load might shed light on the nature of the AB signalling process. The bud transcriptome of de-fruited citrus trees was compared with those of ON- and OFF-Crop trees. Fruit removal resulted in relatively rapid changes in global gene expression, including induction of photosynthetic genes and proteins. Altered regulatory mechanisms included abscisic acid (ABA) metabolism and auxin polar transport. Genes of ABA biosynthesis were induced; however, hormone analyses showed that the ABA level was reduced in OFF-Crop buds and in buds shortly following fruit removal. Additionally, genes associated with Ca(2+)-dependent auxin polar transport were remarkably induced in buds of OFF-Crop and de-fruited trees. Hormone analyses showed that auxin levels were reduced in these buds as compared with ON-Crop buds. In view of the auxin transport autoinhibition theory, the possibility that auxin distribution plays a role in determining bud fate is discussed.

  17. Mum, this bud’s for you: where do you want it? Roles for Cdc42 in controlling bud site selection in Saccharomyces cerevisiae

    OpenAIRE

    Nelson, W. James

    2003-01-01

    The generation of asymmetric cell shapes is a recurring theme in biology. In budding yeast, one form of cell asymmetry occurs for division and is generated by anisotropic growth of the mother cell to form a daughter cell bud. Previous genetic studies uncovered key roles for the small GTPase Cdc42 in organizing the actin cytoskeleton and vesicle delivery to the site of bud growth,(1,2) but a recent paper has also raised questions about how control of Cdc42 activity is integrated into a propose...

  18. Accelerated turnover of taste bud cells in mice deficient for the cyclin-dependent kinase inhibitor p27Kip1

    Directory of Open Access Journals (Sweden)

    Perna Marla K

    2011-04-01

    Full Text Available Abstract Background Mammalian taste buds contain several specialized cell types that coordinately respond to tastants and communicate with sensory nerves. While it has long been appreciated that these cells undergo continual turnover, little is known concerning how adequate numbers of cells are generated and maintained. The cyclin-dependent kinase inhibitor p27Kip1 has been shown to influence cell number in several developing tissues, by coordinating cell cycle exit during cell differentiation. Here, we investigated its involvement in the control of taste cell replacement by examining adult mice with targeted ablation of the p27Kip1 gene. Results Histological and morphometric analyses of fungiform and circumvallate taste buds reveal no structural differences between wild-type and p27Kip1-null mice. However, when examined in functional assays, mutants show substantial proliferative changes. In BrdU incorporation experiments, more S-phase-labeled precursors appear within circumvallate taste buds at 1 day post-injection, the earliest time point examined. After 1 week, twice as many labeled intragemmal cells are present, but numbers return to wild-type levels by 2 weeks. Mutant taste buds also contain more TUNEL-labeled cells and 50% more apoptotic bodies than wild-type controls. In normal mice, p27 Kip1 is evident in a subset of receptor and presynaptic taste cells beginning about 3 days post-injection, correlating with the onset of taste cell maturation. Loss of gene function, however, does not alter the proportions of distinct immunohistochemically-identified cell types. Conclusions p27Kip1 participates in taste cell replacement by regulating the number of precursor cells available for entry into taste buds. This is consistent with a role for the protein in timing cell cycle withdrawal in progenitor cells. The equivalence of mutant and wild-type taste buds with regard to cell number, cell types and general structure contrasts with the hyperplasia

  19. A physiologic role for serotonergic transmission in adult rat taste buds.

    Directory of Open Access Journals (Sweden)

    Luc Jaber

    Full Text Available Of the multiple neurotransmitters and neuropeptides expressed in the mammalian taste bud, serotonin remains both the most studied and least understood. Serotonin is expressed in a subset of taste receptor cells that form synapses with afferent nerve fibers (type III cells and was once thought to be essential to neurotransmission (now understood as purinergic. However, the discovery of the 5-HT1A serotonin receptor in a subset of taste receptor cells paracrine to type III cell suggested a role in cell-to-cell communication during the processing of taste information. Functional data describing this role are lacking. Using anatomical and neurophysiological techniques, this study proposes a modulatory role for serotonin during the processing of taste information. Double labeling immunocytochemical and single cell RT-PCR technique experiments documented that 5-HT1A-expressing cells co-expressed markers for type II cells, cells which express T1R or T2R receptors and release ATP. These cells did not co-express type III cells markers. Neurophysiological recordings from the chorda tympani nerve, which innervates anterior taste buds, were performed prior to and during intravenous injection of a 5-HT1A receptor antagonist. These experiments revealed that serotonin facilitates processing of taste information for tastants representing sweet, sour, salty, and bitter taste qualities. On the other hand, injection of ondansetron, a 5-HT3 receptor antagonist, was without effect. Collectively, these data support the hypothesis that serotonin is a crucial element in a finely-tuned feedback loop involving the 5-HT1A receptor, ATP, and purinoceptors. It is hypothesized that serotonin facilitates gustatory signals by regulating the release of ATP through ATP-release channels possibly through phosphatidylinositol 4,5-bisphosphate resynthesis. By doing so, 5-HT1A activation prevents desensitization of post-synaptic purinergic receptors expressed on afferent nerve fibers

  20. The effects of heat treatment on technological properties in Red-bud maple (Acer trautvetteri Medw.) wood.

    Science.gov (United States)

    Korkut, Süleyman; Kök, M Samil; Korkut, Derya Sevim; Gürleyen, Tuğba

    2008-04-01

    Heat treatment is often used to improve the dimensional stability of wood. In this study, the effects of heat treatment on technological properties of Red-bud maple (Acer trautvetteri Medw.) wood were examined. Samples obtained from Düzce Forest Enterprises, Turkey, were subjected to heat treatment at varying temperatures (120 degrees C, 150 degrees C and 180 degrees C) and for varying durations (2h, 6h and 10h). The technological properties of heat-treated wood samples and control samples were tested. Compression strength parallel to grain, bending strength, modulus of elasticity in bending, janka-hardness, impact bending strength, and tension strength perpendicular to grain were determined. The results showed that technological strength values decreased with increasing treatment temperature and treatment times. Red-bud maple wood could be utilized by using proper heat treatment techniques with minimal losses in strength values in areas where working, and stability such as in window frames, are important factors.

  1. Immunohistochemical expression of budding uninhibited by benzimidazole related 1 in leukoplakia and oral squamous cell carcinoma

    Directory of Open Access Journals (Sweden)

    Taneeru Sravya

    2016-01-01

    Full Text Available Background: Budding uninhibited by benzimidazole related 1 (BUBR1 is an important protein in the mitotic spindle assembly checkpoint. Alterations in expression of BUBR1 have been reported in many premalignant and malignant lesions. Aim: To compare the expression of BUBR1 with respect to the normal mucosa and degree of dysplasia in oral leukoplakia (OL and also with respect to different histopathological grades of oral squamous cell carcinoma (OSCC. Materials and Methods: Neutral buffered formalin-fixed and paraffin-embedded biopsy specimens 30 each of normal, OL and OSCC tissue were included in this study. The expression of BUBR1 was detected using immunohistochemistry (IHC. The scores obtained were subjected to ANOVA test. Results: Significant correlation was found in immunostaining between normal, dysplasia and OSCC groups with a P value of 0.00001. The expression of BUBR1 was significant when compared with different degrees of dysplasia and in different histopathological grades of OSCC with a P value of 0.00001. Conclusion: Higher IHC scores were obtained with increased histopathological grades of OL and OSCC suggesting its role as a prognostic indicator.

  2. CG Methylation Covaries with Differential Gene Expression between Leaf and Floral Bud Tissues of Brachypodium distachyon.

    Directory of Open Access Journals (Sweden)

    Kyria Roessler

    Full Text Available DNA methylation has the potential to influence plant growth and development through its influence on gene expression. To date, however, the evidence from plant systems is mixed as to whether patterns of DNA methylation vary significantly among tissues and, if so, whether these differences affect tissue-specific gene expression. To address these questions, we analyzed both bisulfite sequence (BSseq and transcriptomic sequence data from three biological replicates of two tissues (leaf and floral bud from the model grass species Brachypodium distachyon. Our first goal was to determine whether tissues were more differentiated in DNA methylation than explained by variation among biological replicates. Tissues were more differentiated than biological replicates, but the analysis of replicated data revealed high (>50% false positive rates for the inference of differentially methylated sites (DMSs and differentially methylated regions (DMRs. Comparing methylation to gene expression, we found that differential CG methylation consistently covaried negatively with gene expression, regardless as to whether methylation was within genes, within their promoters or even within their closest transposable element. The relationship between gene expression and either CHG or CHH methylation was less consistent. In total, CG methylation in promoters explained 9% of the variation in tissue-specific expression across genes, suggesting that CG methylation is a minor but appreciable factor in tissue differentiation.

  3. Maintenance of cellular ATP level by caloric restriction correlates chronological survival of budding yeast

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Joon-Seok; Lee, Cheol-Koo, E-mail: cklee2005@korea.ac.kr

    2013-09-13

    Highlights: •CR decreases total ROS and mitochondrial superoxide during the chronological aging. •CR does not affect the levels of oxidative damage on protein and DNA. •CR contributes extension of chronological lifespan by maintenance of ATP level -- Abstract: The free radical theory of aging emphasizes cumulative oxidative damage in the genome and intracellular proteins due to reactive oxygen species (ROS), which is a major cause for aging. Caloric restriction (CR) has been known as a representative treatment that prevents aging; however, its mechanism of action remains elusive. Here, we show that CR extends the chronological lifespan (CLS) of budding yeast by maintaining cellular energy levels. CR reduced the generation of total ROS and mitochondrial superoxide; however, CR did not reduce the oxidative damage in proteins and DNA. Subsequently, calorie-restricted yeast had higher mitochondrial membrane potential (MMP), and it sustained consistent ATP levels during the process of chronological aging. Our results suggest that CR extends the survival of the chronologically aged cells by improving the efficiency of energy metabolism for the maintenance of the ATP level rather than reducing the global oxidative damage of proteins and DNA.

  4. Effect of temperature on replicative aging of the budding yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Molon, Mateusz; Zadrag-Tecza, Renata

    2016-04-01

    The use of the budding yeast Saccharomyces cerevisiae in gerontological studies was based on the assumption that the reproduction limit of a single cell (replicative aging) is a consequence of accumulation of a hypothetical universal "senescence factor" within the mother cell. However, some evidence suggests that molecules or structures proposed as the "aging factor", such as rDNA circles, oxidatively damaged proteins (with carbonyl groups) or mitochondria, have little effect on replicative lifespan of yeast cells. Our results also suggest that protein aggregates associated with Hsp104, treated as a marker of yeast aging, do not seem to affect the numeric value of replicative lifespan of yeast. What these results indicate, however, is the need for finding a different way of expressing age and longevity of yeast cells instead of the commonly used number of daughters produced over units of time, as in the case of other organisms. In this paper, we show that the temperature has a stronger influence on the time of life (the total lifespan) than on the reproductive potential of yeast cells.

  5. Partial purification of histone H3 proteolytic activity from the budding yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Azad, Gajendra Kumar; Tomar, Raghuvir Singh

    2016-06-01

    The proteolytic clipping of histone tails has recently emerged as a novel form of irreversible post-translational modification (PTM) of histones. Histone clipping has been implicated as a regulatory process leading to the permanent removal of PTMs from histone proteins. However, there is scarcity of literature that describes the identification and characterization of histone-specific proteases. Here, we employed various biochemical methods to report histone H3-specific proteolytic activity from budding yeast. Our results demonstrate that H3 proteolytic activity was associated with sepharose bead matrices and activity was not affected by a variety of stress conditions. We have also identified the existence of an unknown protein that acts as a physiological inhibitor of the H3-clipping activity of yeast H3 protease. Moreover, through protease inhibition assays, we have also characterized yeast H3 protease as a serine protease. Interestingly, unlike glutamate dehydrogenase (GDH), yeast H3 proteolytic activity was not inhibited by Stefin B. Together, our findings suggest the existence of a novel H3 protease in yeast that is different from other reported histone H3 proteases. The presence of histone H3 proteolytic activity, along with the physiological inhibitor in yeast, suggests an interesting molecular mechanism that regulates the activity of histone proteases. Copyright © 2016 John Wiley & Sons, Ltd.

  6. Ndc10 is a platform for inner kinetochore assembly in budding yeast

    Energy Technology Data Exchange (ETDEWEB)

    Cho, Uhn-Soo; Harrison, Stephen C. (Harvard-Med)

    2012-01-10

    Kinetochores link centromeric DNA to spindle microtubules and ensure faithful chromosome segregation during mitosis. In point-centromere yeasts, the CBF3 complex Skp1-Ctf13-(Cep3){sub 2}-(Ndc10){sub 2} recognizes a conserved centromeric DNA element through contacts made by Cep3 and Ndc10. We describe here the five-domain organization of Kluyveromyces lactis Ndc10 and the structure at 2.8 {angstrom} resolution of domains I-II (residues 1-402) bound to DNA. The structure resembles tyrosine DNA recombinases, although it lacks both endonuclease and ligase activities. Structural and biochemical data demonstrate that each subunit of the Ndc10 dimer binds a separate fragment of DNA, suggesting that Ndc10 stabilizes a DNA loop at the centromere. We describe in vitro association experiments showing that specific domains of Ndc10 interact with each of the known inner-kinetochore proteins or protein complexes in budding yeast. We propose that Ndc10 provides a central platform for inner-kinetochore assembly.

  7. Direct Adventitious Bud Induction and Plant Regeneration of Rosa hybrida Samantha

    Institute of Scientific and Technical Information of China (English)

    GAO Li-ping; BAO Man-zhu

    2005-01-01

    Effect of explant, site of leaflet, induction period in the dark and combinations of plant growth regulators on direct adventitious bud induction and plant regeneration of Rosa hybrida Samantha was investigated. The results showed that after an induction period of 8 d on MS medium with 1.5 mg L-1 TDZ and 0.05 mg L-1 NAA in the dark and a subculture on MS medium with 0.5 mg L-1 BA and 0.01 mg L-1 NAA under light, the best plant regeneration was obtained and the regeneration frequencies of leaflets and petioles were 51.8 and 10% respectively. There was no significant difference in regeneration ability between leaflets at different sites of the compound leaves, longer time of induction in the dark or high concentration of auxin would cause callus formation, which was disadvantageous for shoot regeneration, and the regeneration frequency was significantly reduced. This regeneration system could be applied for genetic transformation of this cultivar in the future.

  8. FT-IR microscopic mappings of early mineralization in chick limb bud mesenchymal cell cultures

    Science.gov (United States)

    Boskey, A. L.; Camacho, N. P.; Mendelsohn, R.; Doty, S. B.; Binderman, I.

    1992-01-01

    Chick limb bud mesenchymal cells differentiate into chondrocytes and form a cartilaginous matrix in culture. In this study, the mineral formed in different areas within cultures supplemented with 4 mM inorganic phosphate, or 2.5, 5.0, and 10 mM beta-glycerophosphate (beta GP), was characterized by Fourier-transform infrared (FT-IR) microscopy. The relative mineral-to-matrix ratios, and distribution of crystal sizes at specific locations throughout the matrix were measured from day 14 to day 30. The only mineral phase detected was a poorly crystalline apatite. Cultures receiving 4 mM inorganic phosphate had smaller crystals which were less randomly distributed around the cartilage nodules than those in the beta GP-treated cultures. beta GP-induced mineral consisted of larger, more perfect apatite crystals. In cultures receiving 5 or 10 mM beta GP, the relative mineral-to-matrix ratios (calculated from the integrated intensities of the phosphate and amide I bands, respectively) were higher than in the cultures with 4 mM inorganic phosphate or in the in vivo calcified chick cartilage.

  9. Synaptic communication and signal processing among sensory cells in taste buds

    Science.gov (United States)

    Chaudhari, Nirupa

    2014-01-01

    Taste buds (sensory structures embedded in oral epithelium) show a remarkable diversity of transmitters synthesized and secreted locally. The known transmitters accumulate in a cell type selective manner, with 5-HT and noradrenaline being limited to presynaptic cells, GABA being synthesized in both presynaptic and glial-like cells, and acetylcholine and ATP used for signalling by receptor cells. Each of these transmitters participates in local negative or positive feedback circuits that target particular cell types. Overall, the role of ATP is the best elucidated. ATP serves as a principal afferent transmitter, and also is the key trigger for autocrine positive feedback and paracrine circuits that result in potentiation (via adenosine) or inhibition (via GABA or 5-HT). While many of the cellular receptors and mechanisms for these circuits are known, their impact on sensory detection and perception remains to be elaborated in most instances. This brief review examines what is known, and some of the open questions and controversies surrounding the transmitters and circuits of the taste periphery. PMID:24665098

  10. Maintenance of cellular ATP level by caloric restriction correlates chronological survival of budding yeast

    International Nuclear Information System (INIS)

    Highlights: •CR decreases total ROS and mitochondrial superoxide during the chronological aging. •CR does not affect the levels of oxidative damage on protein and DNA. •CR contributes extension of chronological lifespan by maintenance of ATP level -- Abstract: The free radical theory of aging emphasizes cumulative oxidative damage in the genome and intracellular proteins due to reactive oxygen species (ROS), which is a major cause for aging. Caloric restriction (CR) has been known as a representative treatment that prevents aging; however, its mechanism of action remains elusive. Here, we show that CR extends the chronological lifespan (CLS) of budding yeast by maintaining cellular energy levels. CR reduced the generation of total ROS and mitochondrial superoxide; however, CR did not reduce the oxidative damage in proteins and DNA. Subsequently, calorie-restricted yeast had higher mitochondrial membrane potential (MMP), and it sustained consistent ATP levels during the process of chronological aging. Our results suggest that CR extends the survival of the chronologically aged cells by improving the efficiency of energy metabolism for the maintenance of the ATP level rather than reducing the global oxidative damage of proteins and DNA

  11. In Vitro Shoot Bud Differentiation from Hypocotyl Explants of Chili Peppers (Capsicum annuum L.

    Directory of Open Access Journals (Sweden)

    Owk ANIEL KUMAR

    2010-03-01

    Full Text Available Chili pepper (Capsicum annuum L. is an economically important spice crop in tropical and subtropical countries. In vitro plant regeneration was obtained from 15th day old hypocotyl explants of three chili pepper cultivars (Capsicum annuum L., var. �X-235�, var. �PC-1� and var. �Pusa Jwala�. Among the genotypes of Capsicum L. var. �X-235� responded better than the var. �PC-1� and var. �Pusa Jwala�. MS medium containing BAP (4.0 mg/l and IAA (0.5 mg/l was found to be the best medium for the production of maximum number of shoot buds in all the genotypes of chili pepper i.e., 6.80�0.16 (var. �X-235�, 5.00�0.19 (var. �PC-1� and 4.80�0.12 (var. �Pusa Jwala�. The shoots were rooted on MS medium fortified with IBA (0.5 mg/l. Rooted plants were hardened and transplanted to the soil. The plants showed 80-90% survival during transplantation.

  12. CENP-A exceeds microtubule attachment sites in centromere clusters of both budding and fission yeast.

    Science.gov (United States)

    Coffman, Valerie C; Wu, Pengcheng; Parthun, Mark R; Wu, Jian-Qiu

    2011-11-14

    The stoichiometries of kinetochores and their constituent proteins in yeast and vertebrate cells were determined using the histone H3 variant CENP-A, known as Cse4 in budding yeast, as a counting standard. One Cse4-containing nucleosome exists in the centromere (CEN) of each chromosome, so it has been assumed that each anaphase CEN/kinetochore cluster contains 32 Cse4 molecules. We report that anaphase CEN clusters instead contained approximately fourfold more Cse4 in Saccharomyces cerevisiae and ~40-fold more CENP-A (Cnp1) in Schizosaccharomyces pombe than predicted. These results suggest that the number of CENP-A molecules exceeds the number of kinetochore-microtubule (MT) attachment sites on each chromosome and that CENP-A is not the sole determinant of kinetochore assembly sites in either yeast. In addition, we show that fission yeast has enough Dam1-DASH complex for ring formation around attached MTs. The results of this study suggest the need for significant revision of existing CEN/kinetochore architectural models. PMID:22084306

  13. Cell wall and lipid composition of Isosphaera pallida, a budding eubacterium from hot springs.

    Science.gov (United States)

    Giovannoni, S J; Godchaux, W; Schabtach, E; Castenholz, R W

    1987-06-01

    Isosphaera pallida is an unusual gliding, budding eubacterium recently isolated from North American hot springs. Electron micrographs of ultrathin sections revealed a cell wall atypical of eubacteria: two electrondense layers separated by an electron-transparent layer, with no evident peptidoglycan layer. Growth was not inhibited by penicillin. Cell walls were isolated from sheared cells by velocity sedimentation. The rigid-layer fraction, prepared from cell walls by treatment with boiling 10% sodium dodecyl sulfate, was hydrolyzed and chemically analyzed for muramic acid. This essential component of peptidoglycan was absent. Amino acid analysis demonstrated a proteinaceous wall structure. Pitlike surface structures seen in negatively stained whole cells and thin sections were correlated with periodically spaced perforations of the rigid sacculus. An analysis of the lipid composition of I. pallida revealed typical ester-linked lipids with unbranched fatty acids, in contrast to the isoprenyl ether-linked lipids of archaebacteria, which also have proteinaceous cell walls. Capnoids, unusual sulfonolipids which are present in gliding bacteria of the Cytophaga-Flexibacter group, were absent. PMID:3584067

  14. [Neurocristopathies of the human nasofrontal bud. Ethmoidal syndromes (hypo- and hyper-septoethmoidism) (author's transl)].

    Science.gov (United States)

    Couly, G

    1981-01-01

    Knowledge of human brain development biology has increased rapidly over the last decade due to fundamental acquisitions in the fields of organogenesis, fetal development, and post-natal growth. The role of the neural crests during cephalic organogenesis, recently studied by Mme Nicole Le Douarin, has given new biological dimensions to the head : it is an organic complex, globally neural in character, and of encephalofacial territorialization. The cervicocephalic region derives this original segmentation from the migratory property of this contingent of the neural crests. In this perspective, the nasofrontal bud is therefore an embryonic cephalic segment, in which appears the pro-encephalon, the eyes, the olfactory bulb, and the median facial zones (bone, cartilage, teeth). Any morphogenetic anomaly in this original craniofacial segment creates associated malformation of the brain, the base of the skull, and the face (eyes and olfactory bulbs) : the neurocristopathies. In this respect, and from the semiological point of view, the face is the "predicate" of the brain. A new classification of these malformations is proposed, based on the clinical study of the ethmoid, a cartilaginous median organ which is accessible clinically. Epigenesis lasts for 18 months in this organ, which forms a part of the face and the neurocranium, and which is known to play a "motor" morphogenetic role on the osteomembranous face : the ethmoidal syndromes (hypo- and hyper-septoethmoidism). Future reports will discuss the neurocristopathies of the branchial arches. PMID:6944757

  15. STIM1 regulates calcium signaling in taste bud cells and preference for fat in mice

    Science.gov (United States)

    Dramane, Gado; Abdoul-Azize, Souleymane; Hichami, Aziz; VÖgtle, Timo; Akpona, Simon; Chouabe, Christophe; Sadou, Hassimi; Nieswandt, Bernhard; Besnard, Philippe; Khan, Naim Akhtar

    2012-01-01

    Understanding the mechanisms underlying oro-gustatory detection of dietary fat is critical for the prevention and treatment of obesity. The lipid-binding glycoprotein CD36, which is expressed by circumvallate papillae (CVP) of the mouse tongue, has been implicated in oro-gustatory perception of dietary lipids. Here, we demonstrate that stromal interaction molecule 1 (STIM1), a sensor of Ca2+ depletion in the endoplasmic reticulum, mediates fatty acid–induced Ca2+ signaling in the mouse tongue and fat preference. We showed that linoleic acid (LA) induced the production of arachidonic acid (AA) and lysophosphatidylcholine (Lyso-PC) by activating multiple phospholipase A2 isoforms via CD36. This activation triggered Ca2+ influx in CD36-positive taste bud cells (TBCs) purified from mouse CVP. LA also induced the production of Ca2+ influx factor (CIF). STIM1 was found to regulate LA-induced CIF production and the opening of multiple store-operated Ca2+ (SOC) channels. Furthermore, CD36-positive TBCs from Stim1–/– mice failed to release serotonin, and Stim1–/– mice lost the spontaneous preference for fat that was observed in wild-type animals. Our results suggest that fatty acid–induced Ca2+ signaling, regulated by STIM1 via CD36, might be implicated in oro-gustatory perception of dietary lipids and the spontaneous preference for fat. PMID:22546859

  16. Visual screening for localized RNAs in yeast revealed novel RNAs at the bud-tip

    International Nuclear Information System (INIS)

    Several RNAs, including rRNAs, snRNAs, snoRNAs, and some mRNAs, are known to be localized at specific sites in a cell. Although methods have been established to visualize RNAs in a living cell, no large-scale visual screening of localized RNAs has been performed. In this study, we constructed a genomic library in which random genomic fragments were inserted downstream of U1A-tag sequences under a GAL1 promoter. In a living yeast cell, transcribed U1A-tagged RNAs were visualized by U1A-GFP that binds the RNA sequence of the U1A-tag. In this screening, many RNAs showed nuclear signals. Since the nuclear signals of some RNAs were not seen when the U1A-tag was connected to the 3' ends of the RNAs, it is suggested that their nuclear signals correspond to nascent transcripts on GAL1 promoter plasmids. Using this screening method, we successfully identified two novel localized mRNAs, CSR2 and DAL81, which showed bud-tip localization

  17. Evidence for widespread adaptive evolution of gene expression in budding yeast.

    Science.gov (United States)

    Fraser, Hunter B; Moses, Alan M; Schadt, Eric E

    2010-02-16

    Changes in gene expression have been proposed to underlie many, or even most, adaptive differences between species. Despite the increasing acceptance of this view, only a handful of cases of adaptive gene expression evolution have been demonstrated. To address this discrepancy, we introduce a simple test for lineage-specific selection on gene expression. Applying the test to genome-wide gene expression data from the budding yeast Saccharomyces cerevisiae, we find that hundreds of gene expression levels have been subject to lineage-specific selection. Comparing these findings with independent population genetic evidence of selective sweeps suggests that this lineage-specific selection has resulted in recent sweeps at over a hundred genes, most of which led to increased transcript levels. Examination of the implicated genes revealed a specific biochemical pathway--ergosterol biosynthesis--where the expression of multiple genes has been subject to selection for reduced levels. In sum, these results suggest that adaptive evolution of gene expression is common in yeast, that regulatory adaptation can occur at the level of entire pathways, and that similar genome-wide scans may be possible in other species, including humans.

  18. The nuclear exosome is active and important during budding yeast meiosis.

    Directory of Open Access Journals (Sweden)

    Stephen Frenk

    Full Text Available Nuclear RNA degradation pathways are highly conserved across eukaryotes and play important roles in RNA quality control. Key substrates for exosomal degradation include aberrant functional RNAs and cryptic unstable transcripts (CUTs. It has recently been reported that the nuclear exosome is inactivated during meiosis in budding yeast through degradation of the subunit Rrp6, leading to the stabilisation of a subset of meiotic unannotated transcripts (MUTs of unknown function. We have analysed the activity of the nuclear exosome during meiosis by deletion of TRF4, which encodes a key component of the exosome targeting complex TRAMP. We find that TRAMP mutants produce high levels of CUTs during meiosis that are undetectable in wild-type cells, showing that the nuclear exosome remains functional for CUT degradation, and we further report that the meiotic exosome complex contains Rrp6. Indeed Rrp6 over-expression is insufficient to suppress MUT transcripts, showing that the reduced amount of Rrp6 in meiotic cells does not directly cause MUT accumulation. Lack of TRAMP activity stabilises ∼ 1600 CUTs in meiotic cells, which occupy 40% of the binding capacity of the nuclear cap binding complex (CBC. CBC mutants display defects in the formation of meiotic double strand breaks (DSBs, and we see similar defects in TRAMP mutants, suggesting that a key function of the nuclear exosome is to prevent saturation of the CBC complex by CUTs. Together, our results show that the nuclear exosome remains active in meiosis and has an important role in facilitating meiotic recombination.

  19. Dynamics of DNA methylation and Histone H4 acetylation during floral bud differentiation in azalea

    Directory of Open Access Journals (Sweden)

    Valledor Luis

    2010-01-01

    Full Text Available Abstract Background The ability to control the timing of flowering is a key strategy for planning production in ornamental species such as azalea, however it requires a thorough understanding of floral transition. Floral transition is achieved through a complex genetic network and regulated by multiple environmental and endogenous cues. Dynamic changes between chromatin states facilitating or inhibiting DNA transcription regulate the expression of floral induction pathways in response to environmental and developmental signals. DNA methylation and histone modifications are involved in controlling the functional state of chromatin and gene expression. Results The results of this work indicate that epigenetic mechanisms such as DNA methylation and histone H4 acetylation have opposite and particular dynamics during the transition from vegetative to reproductive development in the apical shoots of azalea. Global levels of DNA methylation and histone H4 acetylation as well as immunodetection of 5-mdC and acetylated H4, in addition to a morphological study have permitted the delimitation of four basic phases in the development of the azalea bud and allowed the identification of a stage of epigenetic reprogramming which showed a sharp decrease of whole DNA methylation similar to that is defined in other developmental processes in plants and in mammals. Conclusion The epigenetic control and reorganization of chromatin seem to be decisive for coordinating floral development in azalea. DNA methylation and H4 deacetylation act simultaneously and co-ordinately, restructuring the chromatin and regulating the gene expression during soot apical meristem development and floral differentiation.

  20. Zebrafish gcm2 is required for gill filament budding from pharyngeal ectoderm.

    Science.gov (United States)

    Hogan, Benjamin M; Hunter, Michael P; Oates, Andrew C; Crowhurst, Meredith O; Hall, Nathan E; Heath, Joan K; Prince, Victoria E; Lieschke, Graham J

    2004-12-15

    The pharyngeal arches give rise to multiple organs critical for diverse processes, including the thymus, thyroid and parathyroids. Several molecular regulators of thymus and thyroid organogenesis are strikingly conserved between mammals and zebrafish. However, land animals have parathyroids whereas fish have gills. The murine transcription factor Glial cells missing 2 (Gcm2) is expressed specifically in the parathyroid primordium in the endodermal epithelium of the third pharyngeal pouch, and in both mice and humans is required for normal development of parathyroid glands. The molecular regulation of fish gill organogenesis remains to be described. We report the expression of gcm2 in the zebrafish pharyngeal epithelium and a requirement for Hox group 3 paralogs for gcm2 expression. Strikingly, zebrafish gcm2 is expressed in the ectodermal portion of the pharyngeal epithelium and is required for the development of the gill filament buds, precursors of fish-specific gill filaments. This study identifies yet another role for a GCM gene in embryonic development and indicates a role for gcm2 during the evolution of divergent pharyngeal morphologies.

  1. Structure of constituents isolated from the flower buds of Cananga odorata and their inhibitory effects on aldose reductase.

    Science.gov (United States)

    Matsumoto, Takahiro; Nakamura, Seikou; Fujimoto, Katsuyoshi; Ohta, Tomoe; Ogawa, Keiko; Yoshikawa, Masayuki; Matsuda, Hisashi

    2014-10-01

    Three new terpenoid derivatives, canangaterpenes IV-VI, were isolated from the flower buds of Cananga odorata, cultivated in Thailand, together with eight known flavonoids. The chemical structures of the new compounds were elucidated on the basis of chemical and physicochemical evidence. The inhibitory effects of the isolated compounds on aldose reductase were also investigated. Several terpenoid derivatives and flavonoids were shown to inhibit aldose reductase. PMID:24816646

  2. Mitochondrial quality control during inheritance is associated with lifespan and mother-daughter age asymmetry in budding yeast

    OpenAIRE

    McFaline-Figueroa, José Ricardo; Vevea, Jason; Swayne, Theresa C.; Zhou, Chun; Liu, Christopher; Leung, Galen; Boldogh, Istvan R.; Pon, Liza A.

    2011-01-01

    Fluorescence loss in photobleaching experiments and analysis of mitochondrial function using superoxide and redox potential biosensors revealed that mitochondria within individual yeast cells are physically and functionally distinct. Mitochondria that are retained in mother cells during yeast cell division have significantly lower redox potential and higher superoxide levels compared to mitochondria in buds. Retention of mitochondria with lower redox potential in mother cells occurs to the sa...

  3. Scavenger Activity Evaluation of the Clove Bud Essential Oil (Eugenia caryophyllus) and Eugenol Derivatives Employing ABTS+• Decolorization

    OpenAIRE

    Merchán Arenas, Diego R.; Acevedo, Amner Muñoz; Vargas Méndez, Leonor Y.; Kouznetsov, Vladimir V.

    2011-01-01

    The essential oil (EO) of clove bud dried fruits from Eugenia caryophyllus was obtained by a conventional hydrodistillation process in an excellent yield (11.7 %). Its chemical composition was analyzed by GC-MS, identifying eugenol as a main constituent (60.5%). Four eugenol-like molecules, γ-diisoeugenol, hydroxymethyleugenol, dihydroeugenol and 1,3-dioxanylphenol, were synthesized using eugenol or isoeugenol as initial precursors under green chemistry protocols. To evaluate the possible ant...

  4. Marketingový mix multikina Cinestar v Českých Budějovicích

    OpenAIRE

    Kašparová, Barbora

    2010-01-01

    In its theoretic part this bachelor thesis deals with development of multiplexes, describes current state of multiplex market in the Czech republic, specifies the terms marketing and marketing mix and focuses on marketing in the service sector. In the practical part it focuses on marketing mix of a particular multiplex -- CineStar České Budějovice. The aim of this thesis is to analyse each component of marketing mix, evaluate it, eventually suggest possible solutions.

  5. Effect of light intensity, plant density and flower bud removal on the flower size and number in cut chrysanthemum

    OpenAIRE

    Carvalho, S.M.P.; Heuvelink, E.; Kooten, van, J.

    2002-01-01

    Flower size and number of flowers per plant are important external quality aspects in cut chrysanthemum. The present work is conducted in a glasshouse and aims at investigating how these quality aspects can be predicted. To evaluate individual flower size, different levels of supplementary lighting (control and assimilation light), plant density (32, 48 and 64 plants m-2) and lateral flower bud removal (leaving 1 flower, 4 flowers and control) were applied. To analyse the effect of assimilate...

  6. Assessment of sanitary conditions in stands of Norway spruce (Picea abies Karst. damaged by spruce bud scale (Physokermes piceae Schrnk.

    Directory of Open Access Journals (Sweden)

    Miezite O

    2013-02-01

    Full Text Available Spruce bud scale (Physokermes piceae Schrnk. affects tree growth directly and indirectly. Direct injury appears in the form of tissue damage, as insects suck sap from tree phloem. Indirect injury appears as “honeydew”, which results in negative effects on tree growth. Plant sap is saturated with various carbohydrates called photosynthates that are difficult for scaly insects to digest. Therefore it is secreted in excrements, which are subsequently a food source for the black sooty mold (Apiosporium pinophilum Fuckel. The fungus covers needles blocking stomata, causing decreased transpiration and photosynthesis. An inexplicable wither of Norway spruce was reported in Latvia during 2010 due to black sooty mold. However, spruce bud scale was not evident. In 2011, mass propagation of spruce bud scale was observed following the 2010 Norway spruce loss. One objective of this research was to determine if Kraft tree growth classes could be applied to establish the factors responsible for tree foliage damage. Six 21 - 40 year old (second age class Norway spruce stands were evaluated. Two circular sample plots with a 7.98 m radius, and a 200-m2 area were randomly established per each forest stand hectare. Diameter at breast height (dbh, 1.3 m, and height of approximately 30 trees was measured to model a trend. For all trees, Kraft class, and foliage damage level caused by spruce bud scale and black sooty mold were determined. Significant differences were not observed in tree damage levels among stands, however significant differences among damage levels in different Kraft classes were detected (F = 3.45 > Fcrit. = 2.80, α = 0.05 > P = 0.02 found. Overall damage intensity was 29.3 %. Total forestry loss was 1153 LVL (1640 EUR for all surveyed stands (10 ha, and 115 LVL (164 EUR per hectare.

  7. FLO1 Is a Variable Green Beard Gene that Drives Biofilm-like Cooperation in Budding Yeast

    OpenAIRE

    Smukalla, Scott; Caldara, Marina; Pochet, Nathalie; Beauvais, Anne; Guadagnini, Stephanie; Yan, Chen; Vinces, Marcelo; Jansen, An; Prevost, Marie Christine; Latge, Jean-Paul; Fink, Gerald R.; Foster, Kevin R.; Verstrepen, Kevin

    2008-01-01

    The budding yeast, Saccharomyces cerevisiae, has emerged as an archetype of eukaryotic cell biology. Here we show that S. cerevisiae is also a model for the evolution of cooperative behavior by revisiting flocculation, a self-adherence phenotype lacking in most laboratory strains. Expression of the gene FLO1 in the laboratory strain S288C restores flocculation, an altered physiological state, reminiscent of bacterial biofilms. Flocculation protects the FLO1 expressing cells from multiple stre...

  8. Effects of allspice, cinnamon, and clove bud essential oils in edible apple films on physical properties and antimicrobial activities.

    Science.gov (United States)

    Du, W-X; Olsen, C W; Avena-Bustillos, R J; McHugh, T H; Levin, C E; Friedman, Mendel

    2009-09-01

    Essential oils (EOs) derived from plants are rich sources of volatile terpenoids and phenolic compounds. Such compounds have the potential to inactivate pathogenic bacteria on contact and in the vapor phase. Edible films made from fruits or vegetables containing EOs can be used commercially to protect food against contamination by pathogenic bacteria. EOs from cinnamon, allspice, and clove bud plants are compatible with the sensory characteristics of apple-based edible films. These films could extend product shelf life and reduce risk of pathogen growth on food surfaces. This study evaluated physical properties (water vapor permeability, color, tensile properties) and antimicrobial activities against Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes of allspice, cinnamon, and clove bud oils in apple puree film-forming solutions formulated into edible films at 0.5% to 3% (w/w) concentrations. Antimicrobial activities were determined by 2 independent methods: overlay of the film on top of the bacteria and vapor phase diffusion of the antimicrobial from the film to the bacteria. The antimicrobial activities against the 3 pathogens were in the following order: cinnamon oil > clove bud oil > allspice oil. The antimicrobial films were more effective against L. monocytogenes than against the S. enterica. The oils reduced the viscosity of the apple solutions and increased elongation and darkened the colors of the films. They did not affect water vapor permeability. The results show that apple-based films with allspice, cinnamon, or clove bud oils were active against 3 foodborne pathogens by both direct contact with the bacteria and indirectly by vapors emanating from the films. PMID:19895483

  9. Biomolecules and Natural Medicine Preparations: Analysis of New Sources of Bioactive Compounds from Ribes and Rubus spp. Buds

    Directory of Open Access Journals (Sweden)

    Dario Donno

    2016-02-01

    Full Text Available It is well known that plants are important sources for the preparation of natural remedies as they contain many biologically active compounds. In particular, polyphenols, terpenic compounds, organic acids, and vitamins are the most widely occurring groups of phytochemicals. Some endemic species may be used for the production of herbal preparations containing phytochemicals with significant bioactivity, as antioxidant activity and anti-inflammatory capacities, and health benefits. Blackberry sprouts and blackcurrant buds are known to contain appreciable levels of bioactive compounds, including flavonols, phenolic acids, monoterpenes, vitamin C, and catechins, with several clinical effects. The aim of this research was to perform an analytical study of blackcurrant and blackberry bud-preparations, in order to identify and quantify the main biomarkers, obtaining a specific phytochemical fingerprint to evaluate the single botanical class contribution to total phytocomplex and relative bioactivity, using a High Performance Liquid Chromatograph−Diode Array Detector; the same analyses were performed both on the University laboratory and commercial preparations. Different chromatographic methods were used to determine concentrations of biomolecules in the preparations, allowing for quantification of statistically significant differences in their bioactive compound content both in the case of Ribes nigrum and Rubus cultivated varieties at different harvest stages. In blackcurrant bud-extracts the most important class was organic acids (50.98% followed by monoterpenes (14.05%, while in blackberry preparations the main bioactive classes were catechins (50.06% and organic acids (27.34%. Chemical, pharmaceutical and agronomic-environmental knowledge could be important for obtaining label certifications for the valorization of specific genotypes, with high clinical and pharmaceutical value: this study allowed to develop an effective tool for the natural

  10. Biomolecules and Natural Medicine Preparations: Analysis of New Sources of Bioactive Compounds from Ribes and Rubus spp. Buds.

    Science.gov (United States)

    Donno, Dario; Mellano, Maria Gabriella; Cerutti, Alessandro Kim; Beccaro, Gabriele Loris

    2016-01-01

    It is well known that plants are important sources for the preparation of natural remedies as they contain many biologically active compounds. In particular, polyphenols, terpenic compounds, organic acids, and vitamins are the most widely occurring groups of phytochemicals. Some endemic species may be used for the production of herbal preparations containing phytochemicals with significant bioactivity, as antioxidant activity and anti-inflammatory capacities, and health benefits. Blackberry sprouts and blackcurrant buds are known to contain appreciable levels of bioactive compounds, including flavonols, phenolic acids, monoterpenes, vitamin C, and catechins, with several clinical effects. The aim of this research was to perform an analytical study of blackcurrant and blackberry bud-preparations, in order to identify and quantify the main biomarkers, obtaining a specific phytochemical fingerprint to evaluate the single botanical class contribution to total phytocomplex and relative bioactivity, using a High Performance Liquid Chromatograph-Diode Array Detector; the same analyses were performed both on the University laboratory and commercial preparations. Different chromatographic methods were used to determine concentrations of biomolecules in the preparations, allowing for quantification of statistically significant differences in their bioactive compound content both in the case of Ribes nigrum and Rubus cultivated varieties at different harvest stages. In blackcurrant bud-extracts the most important class was organic acids (50.98%) followed by monoterpenes (14.05%), while in blackberry preparations the main bioactive classes were catechins (50.06%) and organic acids (27.34%). Chemical, pharmaceutical and agronomic-environmental knowledge could be important for obtaining label certifications for the valorization of specific genotypes, with high clinical and pharmaceutical value: this study allowed to develop an effective tool for the natural preparation quality

  11. Human Mesenchymal Stem Cells Derived From Limb Bud Can Differentiate into All Three Embryonic Germ Layers Lineages

    OpenAIRE

    Jiao, Fei; Wang, Juan; Dong, Zhao-lun; Wu, Min-juan; Zhao, Ting-bao; Li, Dan-Dan; Xin WANG

    2012-01-01

    Mesenchymal stem cells (MSCs) have been isolated from many sources, including adults and fetuses. Previous studies have demonstrated that, compared with their adult counterpart, fetal MSCs with several remarkable advantages may be a better resource for clinical applications. In this study, we successfully isolated a rapidly proliferating cell population from limb bud of aborted fetus and termed them “human limb bud–derived mesenchymal stem cells” (hLB-MSCs). Characteristics of their morpholog...

  12. Analýza cestovního ruchu ve vybraném městě (České Budějovice)

    OpenAIRE

    Tupá, Petra

    2009-01-01

    The analysis is concernd with the tourism industry in České Budějovice. The offer of the tourism industry in the city was described, the SWOT analysis is involved too and the future plans are outlined.

  13. The acropetal effects of indole-3-acetic acid in isolated shoot segments of Acer pseudoplatanus L. I. Growth responses of buds

    Directory of Open Access Journals (Sweden)

    Jacek A. Adamczyk

    2014-02-01

    Full Text Available The subject of this investigation were growth reactions of two opposite lateral buds of Acer pseudoplatanus L. stem sections in response to application of indole-3-acetic acid (IAA and other synthetic growth regulators below the buds. No effect of IAA upon the initiation of bud growth was noted, however, elongation of new shoots was inhibited. This acropetal effect of auxin was enhanced by simultaneous treatment with triiodobenzoic acid (TIBA or benzyladenine (BA. TIBA alone caused even stronger retardation of shoot growth. Exactly the opposite effect was produced by gibberellic acid (GA3 applied even 30 cm below the buds. In this case TIBA could reverse the effect of GA3 treatment. The results concerning acropetal effects of auxin suggest that some type of signalling system functions independently of the direct action of the exogenous IAA.

  14. Identification of irradiated food. I.- A test established on the in vitro culture of potato buds to identified the irradiated tubers

    International Nuclear Information System (INIS)

    A method based upon the in vitro culture of potato buds in a mineral medium is described, by which method tubers irradiated can be distinguished from tubers treated by refrigeration or inhibited by chemical agents. (Author) 9 refs

  15. The effect of the time and the budding method on the growth of young cherry trees cv. 'Łutówka'

    Directory of Open Access Journals (Sweden)

    Piotr Baryła

    2012-12-01

    Full Text Available The studies concerning the effect of the time and the methods of budding on the growth of young cherry trees were conducted in the years at Felin Experimental Farm of Lublin Agricultural University. The objects of investigations were the young cherry trees obtained as a result of budding of mahaleb cherry (Prunus mahaleb L. and sweet cherry (Prunus avium L. seedlings in the way by the chip budding-15th July and T-graft-15th July and 1st September. The used methods and the times of budding insignificantly affected the growth of young cherry trees cv. «Łutówka» in a nursery. There was showed that quality features of the trees were dependet on stock used type. Cherry trees obtained on mahaleb cherry were thicker, higher and better branched than on sweet cherry.

  16. Isolation of a cdc28 mutation that abrogates the dependence of S phase on completion of M phase of the budding yeast cell cycle

    Indian Academy of Sciences (India)

    Santanu Kumar Ghosh; Pratima Sinha

    2000-01-01

    We have isolated a mutation in the budding yeast Saccharomyces cerevisisae CDC28 gene that allows cdc13 cells, carrying damaged DNA, to continue with the cell division cycle. While cdc13 mutant cells are arrested as large-budded cells at the nonpermissive temperature 37°C, the cdc13 cdc28 double mutant culture showed cells with one or more buds, most of which showed apical growth. The additional buds emerged without the intervening steps of nuclear division and cell separation. We suggest that the cdc28 mutation abrogates a checkpoint function and allows cells with damaged or incompletely replicated DNA an entry to another round of cell cycle and bypasses the mitotic phase of the cell cycle.

  17. Transient etiolation: protochlorophyll(ide) and chlorophyll forms in differentiating plastids of closed and breaking leaf buds of horse chestnut (Aesculus hippocastanum).

    Science.gov (United States)

    Solymosi, Katalin; Bóka, Károly; Böddi, Béla

    2006-08-01

    An accompanying paper reports the accumulation of photoactive protochlorophyllide (Pchlide) in the innermost leaf primordia of buds of many tree species. In this paper, we describe plastid differentiation, changes in pigment concentrations and spectral properties of bud scales and leaf primordia of horse chestnut (Aesculus hippocastanum L.) from January until the end of bud break in April. The bud scales contained plastids with grana, stroma thylakoids characteristic of chloroplasts and large dense bodies within the stroma. In January, proplastids and young chloroplasts were present in the leaf primordia, and the fluorescence spectra of the primordia were similar to those of green leaves except for a minor band at 630 nm, indicative of a protochlorophyll(ide). During bud break, the pigment concentrations of the green bud scales and the outermost leaf primordia increased, and Pchlide forms with emission maxima at 633, 644 and 655 nm accumulated in the middle and innermost leaf primordia. Depending on the position of the leaf primordia within the bud, their plastids and their pigment concentrations varied. Etio-chloroplasts with prolamellar bodies (PLBs) and prothylakoids with developing grana were observed in the innermost leaves. Besides the above-mentioned Pchlide forms, the middle and innnermost leaf primordia contained only a Chl band with an emission maximum at 686 nm. The outermost leaf primordia contained etio-chloroplasts with well-developed grana and small, narrow-type PLBs. These outermost leaves contained only chlorophyll forms like the mature green leaves. No Pchlide accumulation was observed after bud break, indicating that etiolation of the innermost and middle leaves is transient. The Pchlide forms and the plastid types of the primordia in buds grown in nature were similar to those of leaves of dark-germinated seedlings and to those of the leaf primordia of dark-forced buds. We conclude that transient etiolation occurs under natural conditions. The

  18. Preliminary Observation on Developmental Characteristics of Adventitious Buds of Artemisia frigida Willd.%冷蒿不定芽发育特性的初步观察

    Institute of Scientific and Technical Information of China (English)

    李姗姗; 宛涛; 蔡萍; 伊卫东; 韩轩

    2013-01-01

    对冷蒿不定芽的分布、着生特点、萌生数量及形态结构特征等发育特性的初步观察表明:冷蒿不定芽主要着生于根颈、地表枝条上,其大小、数量以及分布情况各异;不定芽主要产生于返青期到结实期,冷蒿现蕾期和开花期是不定芽萌生的旺盛时期,在根颈处分别有19个和25个,地表枝条上有66个和75个.光镜和扫描电镜观察表明,不定芽具有芽鳞片,包裹着幼叶、叶原基和叶生长点,芽和叶片的表皮被有大量的白色绢毛,发育中的芽体由灰白色转变为绿色,最终发育成叶片和枝条.%The developmental characteristics of adventitious buds of Artemisia frigida Willd. were observed. The results show that the adventitious buds of Artemisia frigida Willd. mainly grow on the rhizome and ground branches,whose size, amount and distribution were different; adventitious buds occurred in May to November, budding stage and flowering stage were period of adventitious buds occurring, there were 19 and 25 adventitious buds on the rhizome and 66 and 75 adventitious buds on the ground branches; by the optical stereoscope and canning electron microscope, the adventitious buds had bud scale, young leaves, leaf primordium and growing tip wrapped in it, the surface of buds and leaves had plenty of white silk wool, the buds turned off-white to green during development, finally formed leaf and branch.

  19. Interaction of environment conditions and genotypes on expression of genetic background in micro-phenophases of strawberry mixed flower bud

    Directory of Open Access Journals (Sweden)

    Selamovska Ana

    2013-01-01

    Full Text Available The aim of this research is differentiation or micro-phenophases of reproductive organs on two junebearing strawberry (Fragaria x anannassa varieties senga sengana and pocahontas, depending on climate conditions, rosettes ordering and cultivate manner (orchard mulched on black foil and orchard on bare soil. The beginning of differentiation of flower buds is genetic characteristic depending on climate conditions (insulations, day length, higher midday and night air temperatures from 1.05 till the beginning of differentiation, the sum of rainfalls from the beginning of May until the end of July, order of rosettes and cultivate manner The sum of effective temperatures over 10ºC from 1st of May till the beginning of differentiation has no influence on beginning of flower buds differentiation. First morphological changes of the apical meristem were started in the first decade of August that has coincided with the day length of 14 hours and day insulations of 9.3 hours. Micro-phenophases were undergoing almost at the same time in both varieties, only the beginning at pocahontas was 2-3 days earlier. Primary rosettes differ 10-15 days earlier than the secondary rosettes. Plants that grown on black foil had 7-10 days earlier flower bud differentiation compared to those grown on bare soil.

  20. Correlation Between Endogenous Hormones of Stem Apices and Fruit Locule Numbers in Tomatoes During Floral Bud Differentiation Stages

    Institute of Scientific and Technical Information of China (English)

    LI Yue; LI Tian-lai; WANG Dan

    2008-01-01

    The study was designed to elucidate the changes in the endogenous hormones of stem apices in tomatoes and the function of correlative endogenous hormones in tomatoes during floral bud differentiation stages. The tomato parents were crossed and reverse crossed by using two inbred lines of multi-locule (MLK1) and few-locule (FL1) with significant difference, and the relationship between endogenous hormones GA3, IAA, and ABA levels and ovary locule numbers of parents and progeny during floral bud differentiation initial stage, floral bud differentiation stage, sepal petal formation stage, carpel formation initial stage, and ovary locule complete formation stage was studied. GA3 levels in P1, P2, F3, and RF1 were consistent with locule numbers, and IAA and ABA levels were reverse to ovary locule numbers during the key stage. The correlation showed that, during sepal petal formation stage, the ovary locule numbers were positively correlated with GA3, GA3/IAA, and GA/ABA, and were negatively correlated with IAA and ABA. It was speculated that increasing GA3 levels or decreasing IAA and ABA levels of stem apices in tomato might be able to enhance ovary locule numbers. The sepal petal formation stage was an important stage which regulated endogenous hormones in the ovary locule formation.

  1. The Effect of Water Stress on Some Morphological, Physiological, and Biochemical Characteristics and Bud Success on Apple and Quince Rootstocks

    Directory of Open Access Journals (Sweden)

    Ibrahim Bolat

    2014-01-01

    Full Text Available The effects of different water stress (control, medium, and severe on some morphological, physiological, and biochemical characteristics and bud success of M9 apple and MA quince rootstocks were determined. The results showed that water stress significantly affected most morphological, physiological, and biochemical characteristics as well as budding success on the both rootstocks. The increasing water stress decreased the relative shoot length, diameter, and plant total fresh and dry weights. Leaf relative water content and chlorophyll index decreased while electrolyte leakage increased with the increase of water stress in both rootstocks. An increase in water stress also resulted in reduction in budding success in Vista Bella/M9 (79.33% and 46.67% and Santa Maria/MA (70.33% and 15.33% combinations. However, the water stress in Santa Maria/MA was more prominent. The increase in water stress resulted in higher peroxidase activities as well as phenol contents in both rootstocks. Although catalase activity, anthocyanin, and proline contents increased with the impact of stress, this was not statistically significant. The results suggest that the impact of stress increased with the increase of water stress; therefore, growers should be careful when using M9 and MA rootstocks in both nursery and orchards where water scarcity is present.

  2. Phenological synchrony between Scaphoideus titanus (Hemiptera: Cicadellidae) hatchings and grapevine bud break: could this explain the insect's expansion?

    Science.gov (United States)

    Chuche, J; Desvignes, E; Bonnard, O; Thiéry, D

    2015-02-01

    Scaphoideus titanus is the invasive vector of the phytoplasma causing the Flavescence dorée in European vineyards. This epidemic is a serious threat to viticulture that has been increasing for more than 60 years in Europe. We studied the effect of synchrony with the plant phenology and the effect of plant-sap quality on the individual fitness. Thus, we conducted laboratory experiments to determine if insect hatchings were synchronized with grapevine bud break. We used two natural populations: one from a cold winter vineyard and one from a mild winter vineyard. In both cases, egg hatching was synchronized with bud break and leaf appearance. The phloem quality of the young and old leaves as a food source was analysed by high-performance liquid chromatography, and the effects on S. titanus growth were evaluated. Phloem composition varied with the grapevine cutting's age but also varied between leaves of different ages from the same plant. The older leaves were less nutritious because they had the highest carbon-to-nitrogen ratio and the lowest content of essential amino acids. Despite diverse phloem qualities, no fitness difference was observed. We found that the synchronization of egg hatchings with bud break is well regulated. However, the nymphs are not affected by the phloem-sap quality, suggesting that S. titanus may accept different food qualities and that egg hatching synchrony could contribute to population expansion in vineyards.

  3. Effects of irradiance on growth and winter bud production by Vallisneria americana and consequences to its abundance and distribution

    Science.gov (United States)

    Korschgen, Carl E.; Green, W.L.; Kenow, Kevin P.

    1997-01-01

    Number, total biomass, and individual mass of winter buds of Vallisneria americana was significantly related to the depth of the 1% of surface irradiance (Z) and the photosynthetic photon irradiance calculated for each shading treatment imposed during this study. Between the range of 23.8 and 111.2 cm depth for the 1% Z, total biomass of winterbuds produced ranged from 0.63 to 0.01 g, counts ranged from 3.5 to 0.1, and mass of individual winterbuds ranged from 0.18 to 0.04 g. Total biomass of winter buds produced was reduced when plants were exposed to a 14-day period without irradiance during the middle of the growing season. Applying the results of the culture experiments to conditions found in Navigation Pool 8 of the Upper Mississippi River suggests that irradiance may indeed limit the distribution and abundance of Vallisneria americana by reducing the number and size of winter buds. (C) 1997 Elsevier Science B.V.

  4. Research on Flower Bud Differentiation of Ardisia crenata%富贵籽花芽分化研究

    Institute of Scientific and Technical Information of China (English)

    黄冬华; 黄长林; 陶秀花; 宋小民; 王华伶; 黄虹; 李宝光

    2012-01-01

    富贵籽花芽由头年生的侧枝顶端发育而成,花芽分化从3月下旬开始至6月上旬完成,历时80 d,花芽分化的过程可划分为:未分化期、开始分化期、花序原基分化期、小花原基分化期、萼片分化期、花瓣分化期、雄蕊分化期和雌蕊分化期8个时期,分化的顺序从下往上、自外向内.%The flower bud of Ardisia crenata developed from the top of lateral branch that lived last year. The flower bud differentiation lasted 80 days from late March to early June. The course of flower bud differentiation could be divided into 8 stages. Un - differentiation , initial differentiation, differentiation of inflorescence anlage, differentiation of floret anlage, differentiation of sepal, differentiation of petal, differentiation of stamen, and differentiation of pistil. The differentiation order was from bottom to top, and from outside to inside.

  5. Crataegus monogyna buds and fruits phenolic extracts: growth inhibitory activity on human tumor cell lines and chemical characterization by HPLC–DAD–ESI/MS

    OpenAIRE

    Rodrigues, Sandra; Ricardo C. Calhelha; Barreira, João C.M.; Dueñas, Montserrat; Carvalho, Ana Maria; Abreu, Rui M.V.; Santos-Buelga, Celestino; Ferreira, Isabel C.F.R.

    2012-01-01

    Crataegus monogyna has been extensively studied due to its various alleged health benefits. This study aimed to determine the human tumor cells growth inhibitory activity of phenolic extracts of its flower buds and fruits in three phenological stages, and further characterize the extracts by HPLC–DAD–ESI/MS. Flower bud extract showed the highest antiproliferative activity as indicated by the lowest GI50 values obtained in all the tested cell lines: MCF-7, breast adenocarcinoma; NC...

  6. Ice barriers promote supercooling and prevent frost injury in reproductive buds, flowers and fruits of alpine dwarf shrubs throughout the summer ☆

    OpenAIRE

    Kuprian, Edith; Briceño, Verónica F.; Wagner, Johanna; Neuner, Gilbert

    2014-01-01

    Over-wintering reproductive buds of many woody plants survive frost by supercooling. The bud tissues are isolated from acropetally advancing ice by the presence of ice barriers that restrict ice growth. Plants living in alpine environments also face the risk of ice formation in summer months. Little knowledge exists, how reproductive structures of woody alpine plants are protected from frost injury during episodic summer frosts. In order to address this question, frost resistance of three com...

  7. In vitro development of buds from tubers of (Solanum tuberosum L.); Desarrollo in vitro de yemas procedentes de tuberculos de patata (Solanum tuberosum L.) tratados por radiacion GAMMA

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez Gonzalez, J.; Garcia Collantes, M. A.

    1976-07-01

    The present work studies the in vitro development of buds from potato tubers subjected to gamma radiation at doses of 3, 6, 9 and 12 Krad. Ths effect of radiation was dependent on the dormant stage of the buds. Intermediate doses (6-9 Krad) did inhibit mitotic division but not cellular elongation. When irradiation is carried out at the end of the resting period, there is an apparent sprouting due to the elongation of previously formed cells. (Author) 17 refs.

  8. Characterization of the minimum domain required for targeting budding yeast myosin II to the site of cell division

    Directory of Open Access Journals (Sweden)

    Tolliday Nicola J

    2006-06-01

    Full Text Available Abstract Background All eukaryotes with the exception of plants use an actomyosin ring to generate a constriction force at the site of cell division (cleavage furrow during mitosis and meiosis. The structure and filament forming abilities located in the C-terminal or tail region of one of the main components, myosin II, are important for localising the molecule to the contractile ring (CR during cytokinesis. However, it remains poorly understood how myosin II is recruited to the site of cell division and how this recruitment relates to myosin filament assembly. Significant conservation between species of the components involved in cytokinesis, including those of the CR, allows the use of easily genetically manipulated organisms, such as budding yeast (Saccharomyces cerevisiae, in the study of cytokinesis. Budding yeast has a single myosin II protein, named Myo1. Unlike most other class II myosins, the tail of Myo1 has an irregular coiled coil. In this report we use molecular genetics, biochemistry and live cell imaging to characterize the minimum localisation domain (MLD of budding yeast Myo1. Results We show that the MLD is a small region in the centre of the tail of Myo1 and that it is both necessary and sufficient for localisation of Myo1 to the yeast bud neck, the pre-determined site of cell division. Hydrodynamic measurements of the MLD, purified from bacteria or yeast, show that it is likely to exist as a trimer. We also examine the importance of a small region of low coiled coil forming probability within the MLD, which we call the hinge region. Removal of the hinge region prevents contraction of the CR. Using fluorescence recovery after photobleaching (FRAP, we show that GFP-tagged MLD is slightly more dynamic than the GFP-tagged full length molecule but less dynamic than the GFP-tagged Myo1 construct lacking the hinge region. Conclusion Our results define the intrinsic determinant for the localization of budding yeast myosin II and show

  9. Dynamic behaviour of the 'Raman spectroscopic signature of life' in a starving budding yeast cell

    International Nuclear Information System (INIS)

    Complete text of publication follows. In vivo molecular-level information is essentially important for understanding the structure, dynamics and functions of living cells. We use Raman microspectroscopy to single living budding yeast cells under a starving condition and study the dynamic behaviour of the 'Raman spectroscopic signature of life' (Y. Naito et al., J. Raman. Spectrosc., 36 (2005) 837-839. and Y-S. Huang et al., Biochemistry, 44 (2005) 10009-10019.) in relation to the formation and disappearance of a 'dancing body' in a vacuole. A focus is placed on the cell death process and the recovery from it. Our previous studies have shown that, under a starving condition, a dancing body appears suddenly in a vacuole and that the 'Raman spectroscopic signature of life' disappears concomitantly indicating the loss of the mitochondrial metabolic activity. This event is followed by gradual deterioration of the cell structure leading to death. This cell death process was visualized at the molecular level by time-resolved Raman imaging. In the present study, we show strong correlations not only between the appearance of a dancing body and the loss of the mitochondrial activity but also between the disappearance of the dancing body and the recovery of the activity. Time- and space-resolved Raman spectra of a single living budding yeast cell were recorded on a confocal Raman microspectrometer with 632.8 nm line of a He-Ne laser for excitation. The laser power at the sample was about 5 mW. The lateral and depth resolutions were about 300 nm with a 100 x oil immersion objective lens (N.A.=1.3) and about 2 μm with a 100 μm pinhole for a confocal detection. Raman spectra were recorded in the wavenumber range 300 - 1800 cm-1 with a spectral resolution of 3 cm-1. The exposure time was 150 sec. Saccharomyces cerevisiae/ Saccharomyces Bayanus hybrid, strain AJL3062 from, was grown at 30 deg C under a stationary cultivation condition in wort medium. The cells dispersed in 1 ml

  10. Analysis of basic leucine zipper genes and their expression during bud dormancy in peach (Prunus persica).

    Science.gov (United States)

    Sun, Ming-Yue; Fu, Xi-Ling; Tan, Qiu-Ping; Liu, Li; Chen, Min; Zhu, Cui-Ying; Li, Ling; Chen, Xiu-De; Gao, Dong-Sheng

    2016-07-01

    Dormancy is a biological characteristic developed to resist the cold conditions in winter. The bZIP transcription factors are present exclusively in eukaryotes and have been identified and classified in many species. bZIP proteins are known to regulate numerous biological processes, however, the role of bZIP in bud dodormancy has not been studied extensively. In total, 50 PpbZIP transcription factor-encoding genes were identified and categorized them into 10 groups (A-I and S). Similar intron/exon structures, additional conserved motifs, and DNA-binding site specificity supported our classification scheme. Additionally, chromosomal distribution and collinearity analyses suggested that expansion of the PpbZIP transcription factor family was due to segment/chromosomal duplications. We also predicted the dimerization properties based on characteristic features of the leucine zipper and classified PpbZIP proteins into 23 subfamilies. Furthermore, qRT-PCR results indicated that PpbZIPs genes may be involved in regulating dormancy. The same gene of different species might participate in different regulating networks through interactions with specific partners. Our expression profiling results complemented the microarray data, suggesting that co-expression patterns of bZIP transcription factors during dormancy differed among deciduous fruit trees. Our findings further clarify the molecular characteristics of the PpbZIP transcription factor family, including potential gene functions during dormancy. This information may facilitate further research on the evolutionary history and biological functions of bZIP proteins in peach and other rosaceae plants. PMID:27107182

  11. Unconventional genomic architecture in the budding yeast saccharomyces cerevisiae masks the nested antisense gene NAG1.

    Science.gov (United States)

    Ma, Jun; Dobry, Craig J; Krysan, Damian J; Kumar, Anuj

    2008-08-01

    The genomic architecture of the budding yeast Saccharomyces cerevisiae is typical of other eukaryotes in that genes are spatially organized into discrete and nonoverlapping units. Inherent in this organizational model is the assumption that protein-coding sequences do not overlap completely. Here, we present evidence to the contrary, defining a previously overlooked yeast gene, NAG1 (for nested antisense gene) nested entirely within the coding sequence of the YGR031W open reading frame in an antisense orientation on the opposite strand. NAG1 encodes a 19-kDa protein, detected by Western blotting of hemagglutinin (HA)-tagged Nag1p with anti-HA antibodies and by beta-galactosidase analysis of a NAG1-lacZ fusion. NAG1 is evolutionarily conserved as a unit with YGR031W in bacteria and fungi. Unlike the YGR031WP protein product, however, which localizes to the mitochondria, Nag1p localizes to the cell periphery, exhibiting properties consistent with those of a plasma membrane protein. Phenotypic analysis of a site-directed mutant (nag1-1) disruptive for NAG1 but silent with respect to YGR031W, defines a role for NAG1 in yeast cell wall biogenesis; microarray profiling of nag1-1 indicates decreased expression of genes contributing to cell wall organization, and the nag1-1 mutant is hypersensitive to the cell wall-perturbing agent calcofluor white. Furthermore, production of Nag1p is dependent upon the presence of the cell wall integrity pathway mitogen-activated protein kinase Slt2p and its downstream transcription factor Rlm1p. Thus, NAG1 is important for two reasons. First, it contributes to yeast cell wall biogenesis. Second, its genomic context is novel, raising the possibility that other nested protein-coding genes may exist in eukaryotic genomes.

  12. Regulation of Budding Yeast CENP-A levels Prevents Misincorporation at Promoter Nucleosomes and Transcriptional Defects.

    Directory of Open Access Journals (Sweden)

    Erica M Hildebrand

    2016-03-01

    Full Text Available The exclusive localization of the histone H3 variant CENP-A to centromeres is essential for accurate chromosome segregation. Ubiquitin-mediated proteolysis helps to ensure that CENP-A does not mislocalize to euchromatin, which can lead to genomic instability. Consistent with this, overexpression of the budding yeast CENP-A(Cse4 is lethal in cells lacking Psh1, the E3 ubiquitin ligase that targets CENP-A(Cse4 for degradation. To identify additional mechanisms that prevent CENP-A(Cse4 misincorporation and lethality, we analyzed the genome-wide mislocalization pattern of overexpressed CENP-A(Cse4 in the presence and absence of Psh1 by chromatin immunoprecipitation followed by high throughput sequencing. We found that ectopic CENP-A(Cse4 is enriched at promoters that contain histone H2A.Z(Htz1 nucleosomes, but that H2A.Z(Htz1 is not required for CENP-A(Cse4 mislocalization. Instead, the INO80 complex, which removes H2A.Z(Htz1 from nucleosomes, promotes the ectopic deposition of CENP-A(Cse4. Transcriptional profiling revealed gene expression changes in the psh1Δ cells overexpressing CENP-A(Cse4. The down-regulated genes are enriched for CENP-A(Cse4 mislocalization to promoters, while the up-regulated genes correlate with those that are also transcriptionally up-regulated in an htz1Δ strain. Together, these data show that regulating centromeric nucleosome localization is not only critical for maintaining centromere function, but also for ensuring accurate promoter function and transcriptional regulation.

  13. Regulation of Budding Yeast CENP-A levels Prevents Misincorporation at Promoter Nucleosomes and Transcriptional Defects.

    Science.gov (United States)

    Hildebrand, Erica M; Biggins, Sue

    2016-03-01

    The exclusive localization of the histone H3 variant CENP-A to centromeres is essential for accurate chromosome segregation. Ubiquitin-mediated proteolysis helps to ensure that CENP-A does not mislocalize to euchromatin, which can lead to genomic instability. Consistent with this, overexpression of the budding yeast CENP-A(Cse4) is lethal in cells lacking Psh1, the E3 ubiquitin ligase that targets CENP-A(Cse4) for degradation. To identify additional mechanisms that prevent CENP-A(Cse4) misincorporation and lethality, we analyzed the genome-wide mislocalization pattern of overexpressed CENP-A(Cse4) in the presence and absence of Psh1 by chromatin immunoprecipitation followed by high throughput sequencing. We found that ectopic CENP-A(Cse4) is enriched at promoters that contain histone H2A.Z(Htz1) nucleosomes, but that H2A.Z(Htz1) is not required for CENP-A(Cse4) mislocalization. Instead, the INO80 complex, which removes H2A.Z(Htz1) from nucleosomes, promotes the ectopic deposition of CENP-A(Cse4). Transcriptional profiling revealed gene expression changes in the psh1Δ cells overexpressing CENP-A(Cse4). The down-regulated genes are enriched for CENP-A(Cse4) mislocalization to promoters, while the up-regulated genes correlate with those that are also transcriptionally up-regulated in an htz1Δ strain. Together, these data show that regulating centromeric nucleosome localization is not only critical for maintaining centromere function, but also for ensuring accurate promoter function and transcriptional regulation.

  14. Alterations of Intracellular Ca2+ Concentration and Ultrastructure in Spruce Apical Bud Cells during Seasonal Transition

    Institute of Scientific and Technical Information of China (English)

    Jian Lingcheng; Sun Delan; Deng Jiangming; Song Yanmei; Paul H. Li

    2004-01-01

    Potassium antimonite was used to localize Ca2+ in the apical bud cells of spruce from July 1999 to May 2000. During the period of active growth (July 14), Calcium precipitates, an indication of Ca2+ localization, were mainly distributed in vacuoles, intercellular spaces and cell walls. Few Ca2+ deposits localized in the cytosol and nucleus, showing a low level of the cytosolic and nuclear Ca2+ concentration in the warm summer. In August, some Ca2+ deposits appeared in the cytosol and nuclei, indicating that Ca2+ influx occurred in the cytosol and nucleus as the day length became shorter. From September to November, high levels of the cytosolic and nuclear Ca2+ remained. During the mid-winter (December and January), the distribution of Ca2+ deposits and the ultrastructures in the cells were altered dramatically. Plasmolysis occurred in many cells due to the protoplasmic dehydration. In addition plasmalemma invagination and nuclear chromatin aggregation also occurred. A large number of Ca2+ deposits appeared in the space between the plasmalemma and the cell wall. And also some Ca2+ deposits were distributed in the plastids. However, few Ca2+ deposits were observed in the cytosol and nuclei. By spring of the next year (May), when plants were de-acclimated and resumed active growth, Ca2+ subcellular localization essentially restored to that observed in July of the last year, i.e., the cells contained low cytosolic and nuclear Ca2+ concentrations; Ca2+ deposits were mainly distributed in the vacuoles, cell walls and intercellular spaces. The relationships between the seasonal changes of intracellular Ca2+ concentration and the development of dormancy/cold acclimation, as well as plasmolysis associated with dormancy and cold hardiness were discussed.

  15. An insight into the complex prion-prion interaction network in the budding yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Du, Zhiqiang; Valtierra, Stephanie; Li, Liming

    2014-01-01

    The budding yeast Saccharomyces cerevisiae is a valuable model system for studying prion-prion interactions as it contains multiple prion proteins. A recent study from our laboratory showed that the existence of Swi1 prion ([SWI(+)]) and overproduction of Swi1 can have strong impacts on the formation of 2 other extensively studied yeast prions, [PSI(+)] and [PIN(+)] ([RNQ(+)]) (Genetics, Vol. 197, 685-700). We showed that a single yeast cell is capable of harboring at least 3 heterologous prion elements and these prions can influence each other's appearance positively and/or negatively. We also showed that during the de novo [PSI(+)] formation process upon Sup35 overproduction, the aggregation patterns of a preexisting inducer ([RNQ(+)] or [SWI(+)]) can undergo significant remodeling from stably transmitted dot-shaped aggregates to aggregates that co-localize with the newly formed Sup35 aggregates that are ring/ribbon/rod- shaped. Such co-localization disappears once the newly formed [PSI(+)] prion stabilizes. Our finding provides strong evidence supporting the "cross-seeding" model for prion-prion interactions and confirms earlier reports that the interactions among different prions and their prion proteins mostly occur at the initiation stages of prionogenesis. Our results also highlight a complex prion interaction network in yeast. We believe that elucidating the mechanism underlying the yeast prion-prion interaction network will not only provide insight into the process of prion de novo generation and propagation in yeast but also shed light on the mechanisms that govern protein misfolding, aggregation, and amyloidogenesis in higher eukaryotes.

  16. Global gene expression analysis of apple fruit development from the floral bud to ripe fruit

    Directory of Open Access Journals (Sweden)

    McArtney Steve

    2008-02-01

    Full Text Available Abstract Background Apple fruit develop over a period of 150 days from anthesis to fully ripe. An array representing approximately 13000 genes (15726 oligonucleotides of 45–55 bases designed from apple ESTs has been used to study gene expression over eight time points during fruit development. This analysis of gene expression lays the groundwork for a molecular understanding of fruit growth and development in apple. Results Using ANOVA analysis of the microarray data, 1955 genes showed significant changes in expression over this time course. Expression of genes is coordinated with four major patterns of expression observed: high in floral buds; high during cell division; high when starch levels and cell expansion rates peak; and high during ripening. Functional analysis associated cell cycle genes with early fruit development and three core cell cycle genes are significantly up-regulated in the early stages of fruit development. Starch metabolic genes were associated with changes in starch levels during fruit development. Comparison with microarrays of ethylene-treated apple fruit identified a group of ethylene induced genes also induced in normal fruit ripening. Comparison with fruit development microarrays in tomato has been used to identify 16 genes for which expression patterns are similar in apple and tomato and these genes may play fundamental roles in fruit development. The early phase of cell division and tissue specification that occurs in the first 35 days after pollination has been associated with up-regulation of a cluster of genes that includes core cell cycle genes. Conclusion Gene expression in apple fruit is coordinated with specific developmental stages. The array results are reproducible and comparisons with experiments in other species has been used to identify genes that may play a fundamental role in fruit development.

  17. Clove bud oil reduces kynurenine and inhibits pqs A gene expression in P. aeruginosa.

    Science.gov (United States)

    H, Jayalekshmi; Omanakuttan, Athira; Pandurangan, N; S Vargis, Vidhu; Maneesh, M; G Nair, Bipin; B Kumar, Geetha

    2016-04-01

    Quorum sensing (QS), a communication system involved in virulence of pathogenic bacteria like Pseudomonas aeruginosa is a promising target to combat multiple drug resistance. In vitro studies using clove bud oil (CBO) in P. aeruginosa revealed a concentration dependent attenuation of a variety of virulence factors including motility, extracellular DNA, exopolysaccharides and pigment production. Furthermore, treatment with CBO demonstrated a distinct dose-dependent reduction in biofilm formation as well as promoting dispersion of already formed biofilm, observations that were also supported by porcine skin ex vivo studies. Expression studies of genes involved in signalling systems of P. aeruginosa indicated a specific decrease in transcription of pqsA, but not in the lasI or rhlI levels. Additionally, the expression of vfr and gacA genes, involved in regulation, was also not affected by CBO treatment. CBO also influenced the PQS signalling pathway by decreasing the levels of kynurenine, an effect which was reversed by the addition of exogenous kynurenine. Though the synthesis of the signalling molecules of the Las and Rhl pathways was not affected by CBO, their activity was significantly affected, as observed by decrease in levels of their various effectors. Molecular modelling studies demonstrated that eugenol, the major component of CBO, favourably binds to the QS receptor by hydrophobic interactions as well as by hydrogen bonding with Arg61 and Tyr41 which are key amino acid residues of the LasR receptor. These results thus elucidate the molecular mechanism underlying the action of CBO and provide the basis for the identification of an attractive QS inhibitor. PMID:26821927

  18. Spatial distribution patterns and environmental interpretation of Anthriscus sylvestris clonal buds

    Institute of Scientific and Technical Information of China (English)

    Weicheng LI; Haiyan SHENG

    2008-01-01

    Anthriscus sylvestris, a weed found both in Europe and China, is a kind of representative clustered clonal plant and is a foe on dams and banks. It has been widely investigated in Europe for its powerful progenitive ability and tolerance to severely adverse environments. Our aims were to investigate and quantify its spatial dis-tribution patterns in four types of community habitats, using a clustering method and adjacency lattice estab-lished by Greig-Smith. We concluded our environmental interpretation based on canonical correspondence ana-lysis (CCA) appended to a Monte Carlo test with rando-mized seeding. The results indicate that the buds around the parent roots are in an aggregation distribution pattern in all scales (0.002-5.12 m2), but theoretical distribution fitting, like negative binomial and Poisson distribution, show that some sizes of several sampling locations are out of place. For this, spatial ordination gives a satisfact-ory answer implying the effect of environmental variables such as depth of humus layer, soil moisture, light con-dition, disturbance intensity and herb abundance. CCA accounts for 64.7% of the total environmental variation and the remaining variation may be counteracted in those five variances or can be interpreted by other factors like accumulating temperature, annual rainfall and altitude in landscape scale. With the aid of temporal sequencing, the suppressed type Ⅱ (monodominant) may be the former mode of suppressed type Ⅰ (stable type), where invasion is done with the help of disturbance from both humans and nature. The abundance of A. sylvestris can add to our cognition in diversity resistance hypothesis and our hypo-thesis on disturbance before or upon immigration.

  19. Comparative live-cell imaging analyses of SPA-2, BUD-6 and BNI-1 in Neurospora crassa reveal novel features of the filamentous fungal polarisome.

    Directory of Open Access Journals (Sweden)

    Alexander Lichius

    Full Text Available A key multiprotein complex involved in regulating the actin cytoskeleton and secretory machinery required for polarized growth in fungi, is the polarisome. Recognized core constituents in budding yeast are the proteins Spa2, Pea2, Aip3/Bud6, and the key effector Bni1. Multicellular fungi display a more complex polarized morphogenesis than yeasts, suggesting that the filamentous fungal polarisome might fulfill additional functions. In this study, we compared the subcellular organization and dynamics of the putative polarisome components BUD-6 and BNI-1 with those of the bona fide polarisome marker SPA-2 at various developmental stages of Neurospora crassa. All three proteins exhibited a yeast-like polarisome configuration during polarized germ tube growth, cell fusion, septal pore plugging and tip repolarization. However, the localization patterns of all three proteins showed spatiotemporally distinct characteristics during the establishment of new polar axes, septum formation and cytokinesis, and maintained hyphal tip growth. Most notably, in vegetative hyphal tips BUD-6 accumulated as a subapical cloud excluded from the Spitzenkörper (Spk, whereas BNI-1 and SPA-2 partially colocalized with the Spk and the tip apex. Novel roles during septal plugging and cytokinesis, connected to the reinitiation of tip growth upon physical injury and conidial maturation, were identified for BUD-6 and BNI-1, respectively. Phenotypic analyses of gene deletion mutants revealed additional functions for BUD-6 and BNI-1 in cell fusion regulation, and the maintenance of Spk integrity. Considered together, our findings reveal novel polarisome-independent functions of BUD-6 and BNI-1 in Neurospora, but also suggest that all three proteins cooperate at plugged septal pores, and their complex arrangement within the apical dome of mature hypha might represent a novel aspect of filamentous fungal polarisome architecture.

  20. 莲藕顶芽繁殖育苗技术应用效果研究%Application Effects of Apical Bud Propagation Technology of Lotus Root

    Institute of Scientific and Technical Information of China (English)

    常发杰; 张锋; 王开昌

    2013-01-01

      为提升保障莲藕供给能力,节本增效,开展莲藕顶芽繁殖和整藕繁殖对产量和效益影响的对比试验。试验结果表明,自莲藕顶芽下1 cm处切下,经杀菌消毒后,移至育苗床,3周后移栽大田,通过施用有机肥和精细田间管理,其效益明显高于整身种藕常规栽培,可大幅降低种子成本,且主藕顶芽繁殖比子藕顶芽繁殖产量更高,具有较大的推广价值。%  In the paper, we studied the effects of the apical bud propagation and conventional lotus root propagation on yield and benefit of lotus root production. We cut the apical buds of lotus root 1 cm under the buds, and moved the buds to the seedling bed after sterilization and disinfection, and then transplanted the seedlings to field three weeks later. The results showed that, the yield and benefit of the apical bud propagation were obviously higher than those of the conventional lotus root propagation by applying organic fertilizers and strengthening field management, and the apical bud propagation method sharply reduced the seed cost of farmers. In addition, the propagation technology using apical buds of mother lotus root possessed higher yield than that of lateral lotus root, and the propagation technology was more worthy of wide promotion.

  1. Digital gene expression analysis of male and female bud transition in Metasequoia reveals high activity of MADS-box transcription factors and hormone-mediated sugar pathways

    Directory of Open Access Journals (Sweden)

    Ying eZhao

    2015-06-01

    Full Text Available Metasequoiaglyptostroboidies is a famous redwood tree of ecological and economic importance, and requires more than 20 years of juvenile-to-adult transition before producing female and male cones. Previously, we induced reproductive buds using a hormone solution in juvenile Metasequoia trees as young as5-to-7years old. In the current study, hormone-treated shoots found in female and male buds were used to identify candidate genes involved in reproductive bud transition in Metasequoia. Samples from hormone-treated cone reproductive shoots and naturally occurring non-cone setting shoots were analyzed using 24 digital gene expression (DGE tag profiles using Illumina, generating a total of 69,520 putative transcripts. Next, 32 differentially and specifically expressed transcripts were determined using quantitative real-time polymerase chain reaction, including the upregulation of MADS-box transcription factors involved in male bud transition and flowering time control proteins involved in female bud transition. These differentially expressed transcripts were associated with 243 KEGG pathways. Among the significantly changed pathways, sugar pathways were mediated by hormone signals during the vegetative-to-reproductive phase transition, including glycolysis/gluconeogenesis and sucrose and starch metabolism pathways. Key enzymes were identified in these pathways, including alcohol dehydrogenase (NAD and glutathione dehydrogenase for the glycolysis/gluconeogenesis pathway, and glucanphosphorylase for sucrose and starch metabolism pathways. Our results increase our understanding of the reproductive bud transition in gymnosperms. In addition, these studies on hormone-mediated sugar pathways increase our understanding of the relationship between sugar and hormone signaling during female and male bud initiation in Metasequoia.

  2. FORECASTING OF GRAPE YIELD AND THE ESTABLISHMENT OF OPTIMUM BUSH LOADING DURING THE CUTTING IN BUDS ON THE PROPOSED YIELD ON THE EXAMPLE OF OAO AF «SOUTH»

    Directory of Open Access Journals (Sweden)

    Matuzok N. V.

    2016-02-01

    Full Text Available The article presents the material of forecasting for grape yield of next year and establishing the optimal loading if cutting of bushes. The material includes 14 varieties of grapes, 11 of them are technical and 3 are table ones. For each year of stable high yield of grapes, it is necessary to pre-set the optimum length of fruit cutting of shoots and optimum load on the bush healthy eyes. To do this for each variety on the eve of trimming bushes we perform optimum productivity analysis of wintering buds of fruit along the length of shoots, i.e. we implement forecasting of grape yield for next year. We have a plan of forecasting for yields of vineyards by microscopy of wintering buds on one-year shoots of fruit ripened grapes in order to establish the potential of embryonic establishment of inflorescences in the central holes of buds. Based on the analysis of buds, the indices were calculated for wintering fruiting buds and their degree of damage during the growing season. It was revealed, that the majority of grape varieties under study shows high tab embryonic inflorescences in central buds in overwintering buds for next year yield. Higher rates at a rate of fruiting buds were wintering in the varieties: Moldova (section 27. - 1.66; Bianca (section 6. - 1.83; Kunlean (section 15. - 1.71; Merlot (section 14. - 1.64; Saperavi (section 56. - 1.76. The lowest rates of fructification - the varieties Muscat Hamburg (section 21 and Augustine (section 11 and were respectively 1.20 and 1.24. As a planned productivity, we offered the optimal loading model of cutting bushes buds. As a result of productivity analyzes of buds along the length of the fruit shoots in 2016 we recommended to carry out pruning of fruit annual shoots 3-4 buds of the form of AZOS-1 and the form of cordon - 5-6 buds

  3. Genetic variation of the bud and leaf phenology of seventeen poplar clones in a short rotation coppice culture.

    Science.gov (United States)

    Pellis, A; Laureysens, I; Ceulemans, R

    2004-01-01

    Leaf phenology of 17 poplar ( Populus spp.) clones, encompassing spring phenology, length of growth period and end-of-year phenology, was examined over several years of different rotations. The 17 poplar clones differed in their latitude of origin (45 degrees 30'N to 51 degrees N) and were studied on a short rotation experimental field plantation, situated in Boom (province of Antwerpen, Belgium; 51 degrees 05'N, 04 degrees 22'E). A similar, clear pattern of bud burst was observed during the different years of study for all clones. Clones Columbia River, Fritzi Pauley, Trichobel (Populus trichocarpa) and Balsam Spire (Populus trichocarpa x Populus balsamifera) from 45 degrees 30'N to 49 degrees N reached bud burst (expressed as day of the year or degree day sums) almost every year earlier than clones Wolterson (Populus nigra), Gaver, Gibecq and Primo (Populus deltoides x Populus nigra) (50 degrees N to 51 degrees N). This observation could not be generalised to end-of-season phenology, for which a yearly returning pattern for all clones was lacking. Late bud burst and early leaf fall of some clones (Beaupré, Boelare, IBW1, IBW2, IBW3) was brought about by increasing rust incidence during the years of observation. For these clones, the variability in leaf phenology was reflected in high coefficients of variation among years. The patterns of genetic variation in leaf phenology have implications for short rotation intensive culture forestry and management of natural populations. Moreover, the variation in phenology reported here is relevant with regard to the genetic mapping of poplar.

  4. Floral bud damage compensation by branching and biomass allocation in genotypes of Brassica napus with different architecture and branching potential.

    Directory of Open Access Journals (Sweden)

    Amélie ePinet

    2015-02-01

    Full Text Available Plant branching is a key process in the yield elaboration of winter oilseed rape (WOSR. It is also involved in plant tolerance to flower damage because it allows the setting of new fertile inflorescences. Here we characterize the changes in the branching and distribution of the number of pods between primary and secondary inflorescences in response to floral bud clippings. Then we investigate the impacts of the modifications in branching on the biomass allocation and its consequence on the crop productivity (harvest index. These issues were addressed on plants with contrasted architecture and branching potential, using three genotypes (Exocet, Pollen, and Gamin grown under two levels of nitrogen fertilization. Clipping treatments of increasing intensities were applied to either inflorescences or flower buds.We were able to show that restoration of the number of pods after clipping is the main lever for the compensation. Genotypes presented different behaviors in branching and biomass allocation as a function of clipping treatments. The number of fertile ramifications increased for the high intensities of clipping. In particular, the growth of secondary ramifications carried by branches developed before clipping has been observed. The proportions of yield and of number of pods carried by these secondary axes increased and became almost equivalent to the proportion carried by primary inflorescences. In terms of biomass allocation, variations have also been evidenced in the relationship between pod dry mass on a given axis and the number of pods set, while the shoot/root ratio was not modified. The harvest index presented different responses: it decreased after flower buds clipping, while it was maintained after the clipping of the whole inflorescences. The results are discussed relative to their implications regarding the identification of interesting traits to be target in breeding programs in order to improve WOSR tolerance.

  5. Novel E3 ubiquitin ligases that regulate histone protein levels in the budding yeast Saccharomyces cerevisiae.

    Directory of Open Access Journals (Sweden)

    Rakesh Kumar Singh

    Full Text Available Core histone proteins are essential for packaging the genomic DNA into chromatin in all eukaryotes. Since multiple genes encode these histone proteins, there is potential for generating more histones than what is required for chromatin assembly. The positively charged histones have a very high affinity for negatively charged molecules such as DNA, and any excess of histone proteins results in deleterious effects on genomic stability and cell viability. Hence, histone levels are known to be tightly regulated via transcriptional, posttranscriptional and posttranslational mechanisms. We have previously elucidated the posttranslational regulation of histone protein levels by the ubiquitin-proteasome pathway involving the E2 ubiquitin conjugating enzymes Ubc4/5 and the HECT (Homologous to E6-AP C-Terminus domain containing E3 ligase Tom1 in the budding yeast. Here we report the identification of four additional E3 ligases containing the RING (Really Interesting New Gene finger domains that are involved in the ubiquitylation and subsequent degradation of excess histones in yeast. These E3 ligases are Pep5, Snt2 as well as two previously uncharacterized Open Reading Frames (ORFs YKR017C and YDR266C that we have named Hel1 and Hel2 (for Histone E3 Ligases respectively. Mutants lacking these E3 ligases are sensitive to histone overexpression as they fail to degrade excess histones and accumulate high levels of endogenous histones on histone chaperones. Co-immunoprecipitation assays showed that these E3 ligases interact with the major E2 enzyme Ubc4 that is involved in the degradation related ubiquitylation of histones. Using mutagenesis we further demonstrate that the RING domains of Hel1, Hel2 and Snt2 are required for histone regulation. Lastly, mutants corresponding to Hel1, Hel2 and Pep5 are sensitive to replication inhibitors. Overall, our results highlight the importance of posttranslational histone regulatory mechanisms that employ multiple E3

  6. Characterization of bud emergence 46 (BEM46) protein: Sequence, structural, phylogenetic and subcellular localization analyses

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Abhishek; Kollath-Leiß, Krisztina; Kempken, Frank, E-mail: fkempken@bot.uni-kiel.de

    2013-08-30

    Highlights: •All eukaryotes have at least a single copy of a bem46 ortholog. •The catalytic triad of BEM46 is illustrated using sequence and structural analysis. •We identified indels in the conserved domain of BEM46 protein. •Localization studies of BEM46 protein were carried out using GFP-fusion tagging. -- Abstract: The bud emergence 46 (BEM46) protein from Neurospora crassa belongs to the α/β-hydrolase superfamily. Recently, we have reported that the BEM46 protein is localized in the perinuclear ER and also forms spots close by the plasma membrane. The protein appears to be required for cell type-specific polarity formation in N. crassa. Furthermore, initial studies suggested that the BEM46 amino acid sequence is conserved in eukaryotes and is considered to be one of the widespread conserved “known unknown” eukaryotic genes. This warrants for a comprehensive phylogenetic analysis of this superfamily to unravel origin and molecular evolution of these genes in different eukaryotes. Herein, we observe that all eukaryotes have at least a single copy of a bem46 ortholog. Upon scanning of these proteins in various genomes, we find that there are expansions leading into several paralogs in vertebrates. Usingcomparative genomic analyses, we identified insertion/deletions (indels) in the conserved domain of BEM46 protein, which allow to differentiate fungal classes such as ascomycetes from basidiomycetes. We also find that exonic indels are able to differentiate BEM46 homologs of different eukaryotic lineage. Furthermore, we unravel that BEM46 protein from N. crassa possess a novel endoplasmic-retention signal (PEKK) using GFP-fusion tagging experiments. We propose that three residues namely a serine 188S, a histidine 292H and an aspartic acid 262D are most critical residues, forming a catalytic triad in BEM46 protein from N. crassa. We carried out a comprehensive study on bem46 genes from a molecular evolution perspective with combination of functional

  7. Generelle aspekter ved mediereception? – Et bud på en multidimensional model for analyse af kvalitative receptionsinterviews

    Directory of Open Access Journals (Sweden)

    Kim Schrøder

    2003-09-01

    Full Text Available Findes der generelle aspekter ved receptionen af medieprodukter, som det kan være analytisk frugtbart at orientere sig efter, og som man altid bør belyse, når man analyserer kvalitative receptionsdata – og måske også allerede når man skal planlægge det empiriske feltarbejde i et em- pirisk receptionsprojekt? Denne artikel bygger på, at dette spørgsmål kan besvares bekræftende, og fremlægger et bud på, hvordan en multi- dimensional model for kvalitativ receptionsanalyse kunne se ud.

  8. Effects of Various Daylength-Treatments on the Growth of Shoots and the Dormancy of Lateral Buds in Mulberry

    OpenAIRE

    YAHIRO, Masaki; YASUHIRO, Norihiko; HIRA, Shigeru; ヤヒロ, マサキ; ヤスヒロ, ノリヒコ; ヒラ, シゲル; 八尋, 正樹; 安広, 紀彦; 平, 茂

    1986-01-01

    In the daylength-treatment of 12 hours, the growth of shoots in the treatment-plot was superior to that in the control-plot (natural daylength-plot; and about 14 hours). In the daylength-treatment of 10 hours, the shoot-growth between the treatment-plot and the control-plot was nearly the same. In the daylength-treatment of 8 hours, the growth of the shoots in the treatment-plot was worse than that in the control-plot. Also, no dormancy of the buds was observed in the daylength-treat...

  9. Effects of Various Daylength-Treatments on the Growth of Shoots and the Dormancy of Lateral Buds in Mulberry

    OpenAIRE

    YAHIRO, Masaki; YASUHIRO, Norihiko; HIRA, Shigeru; ヤヒロ, マサキ; ヤスヒロ, ノリヒコ; ヒラ, シゲル; 八尋, 正樹; 安広, 紀彦; 平, 茂

    1986-01-01

    In the daylength-treatment of 12 hours, the growth of shoots in the treatment-plotwas superior to that in the control-plot (natural daylength-plot; and about14 hours). In the daylength-treatment of 10 hours, the shoot-growth between thetreatment-plot and the control-plot was nearly the same. In the daylength-treatment of 8 hours, the growth of the shoots in the treatment-plot was worse than that in the control-plot. Also, no dormancy of the buds was observed in the daylength-treatments of 8, ...

  10. Induction of shoot buds, multiplication and plantlet formation in seedling explants of bell pepper (Capsicum annuum L. cv. Bryza in vitro

    Directory of Open Access Journals (Sweden)

    Andrzej Gatz

    2014-02-01

    Full Text Available In vitro shoot bud induction and multiplication as well as plantlets formation from different parts of 21-d old seedlings (shoot tip, cotyledonary node, distal part of cotyledon, acropetal section of hypocotyl of Capsicum annuum L., cv. Bryza were compared. During 4 weeks of primary explant culture on initiation media, first shoot bud primordia appeared; they reminded leaf primordia and subsequently some of them underwent enlargement, some developed into leaves and leaf-like structures (mainly on cotyledon explants. The highest number of shoot bud primordia was noted on cotyledonary node explants, but they were smaller than those on the remaining types of the explants. The best response of shoot regeneration showed cotyledon explants on which most of shoot buds were formed in each from four treated passages. From shoot buds on elongation media after 4 weeks of culture rooted rosettes of leaves were achieved, and the extension of the culture time to eight weeks without subculture caused that the rosettes developed into plantlets. Throughout four successive passages plantlets were obtained from cotyledon and shoot tip explants.

  11. Apoplastic H2 O2 plays a critical role in axillary bud outgrowth by altering auxin and cytokinin homeostasis in tomato plants.

    Science.gov (United States)

    Chen, Xiao-Juan; Xia, Xiao-Jian; Guo, Xie; Zhou, Yan-Hong; Shi, Kai; Zhou, Jie; Yu, Jing-Quan

    2016-09-01

    Although phytohormones such as indole-3-acetic acid (IAA), cytokinin (CK) and strigolactone are important modulators of plant architecture, it remains unclear whether reactive oxygen species are involved in the regulation of phytohormone-dependent axillary bud outgrowth in plants. We used diverse techniques, including transcriptional suppression, HPLC-MS, biochemical methodologies and gene transcript analysis to investigate the signaling pathway for apoplastic hydrogen peroxide (H2 O2 )-induced axillary bud outgrowth. Silencing of tomato RESPIRATORY BURST OXIDASE HOMOLOG 1 (RBOH1) and WHITEFLY INDUCED 1 (WFI1), two important genes involved in H2 O2 production in the apoplast, enhanced bud outgrowth, decreased transcript of FZY - a rate-limiting gene in IAA biosynthesis and IAA accumulation in the apex - and increased the transcript of IPT2 involved in CK biosynthesis and CK accumulation in the stem node. These effects were fully abolished by the application of exogenous H2 O2 . Both decapitation and the silencing of FZY promoted bud outgrowth, and downregulated and upregulated the transcripts for IAA3 and IAA15, and IPT2, respectively. However, these effects were not blocked by treatment with exogenous H2 O2 but by napthaleneacetic acid (NAA) treatment. These results suggest that RBOHs-dependent apoplastic H2 O2 promotes IAA biosynthesis in the apex, which, in turn, inhibits CK biosynthesis and subsequent bud outgrowth in tomato plants. PMID:27240824

  12. The Effects of Bud Load and Applied Water Amounts on the Biochemical Composition of the ‘Narince’ Grape Variety (Vitis vinifera L.

    Directory of Open Access Journals (Sweden)

    Serpil GOK TANGOLAR

    2015-12-01

    Full Text Available This study evaluated the effects of various bud load and water amounts (WA on the berry composition of the ‘Narince’ white wine grape variety. Two water amounts (WA-I and WA-II administered during growth stages or a non-irrigated (rain-fed control treatment were paired with two different bud loads (K (normal and 2K (two-fold buds to assess their effects on sugar, organic acid and  phenolic compound contents, as well as antioxidant capacity of the berries for a white wine grape variety ‘Narince’. Despite a slight decrease observed only in sugars in the second year, increases in all phenolic compounds examined (especially in catechin and epicatechin were detected in the WA-I application. The total phenolic compound values obtained for 2K in the same application was also high. The antioxidant capacity values were not significantly affected by the irrigation or bud load applications. Therefore, a 2K bud load with WA-I treatments in which 50% and 75% of the cumulative evaporation from the Class A pan during berry set to veraison and veraison to ripening, respectively, are recommended for irrigation in high plateau viticulture. By maintaining or increasing the fruit composition of ‘Narince’ grapes, these treatments can optimize grape yield and the earnings of growers. In all applications, glucose (among the sugars, tartaric acid (among the organic acids, and catechin and epicatechin (among the phenolic compounds were higher than their counterparts.

  13. Headspace Hanging Drop Liquid Phase Microextraction and Gas Chromatography-Mass Spectrometry for the Analysis of Flavors from Clove Buds

    Energy Technology Data Exchange (ETDEWEB)

    Jung, Mi Jin; Shin, Yeon Jae; Oh, Se Yeon; Kim, Nam Sun; Kim, Kun; Lee, Dong Sun [Seoul Women' s University, Seoul (Korea, Republic of)

    2006-02-15

    A novel sample pretreatment technique, headspace hanging drop liquid phase microextraction (HS-LPME) was studied and applied to the determination of flavors from solid clove buds by gas chromatography-mass spectrometry (GC-MS). Several parameters affecting on HS-LPME such as organic solvent drop volume, extraction time, extraction temperature and phase ratio were investigated. 1-Octanol was selected as the extracting solvent, drop size was fixed to 0.6 μL. 60 min extraction time at 25 .deg. C was chosen. HS-LPME has the good efficiency demonstrated by the higher partition equilibrium constant (K{sub lh}) values and concentration factor (CF) values. The limits of detection (LOD) were 1.5-3.2 ng. The amounts of eugenol, β-caryophyllene and eugenol acetate from the clove bud sample were 1.90 mg/g, 1.47 mg/g and 7.0 mg/g, respectively. This hanging drop based method is a simple, fast and easy sample enrichment technique using minimal solvent. HSLPME is an alternative sample preparation method for the analysis of volatile aroma compounds by GC-MS.

  14. Extraction of primary canine tooth buds: prevalence and associated dental abnormalities in a group of Ethiopian Jewish children.

    Science.gov (United States)

    Holan, G; Mamber, E

    1994-03-01

    Recent publications have described a common belief, held in rural areas in Africa, that unerupted primary canines cause diarrhoea, vomiting and fever in infants. To relieve these symptoms a traditional native healer extracts these tooth buds. The emigration of Ethiopian Jews to Israel in 1991 allowed an investigation of this practice among this community. A group of 59 children (27 boys and 32 girls) aged 3-12 years were examined clinically and radiographically. Evidence was found of extraction of 63 primary canine buds in 35 (59%) of the children. Extraction of one canine was found in 16 children, two in 13 children and three in three children, and three children had all four canines extracted. Forty-six (74%) mandibular compared to only 16 (26%) maxillary canines had been extracted; the extractions were equally divided between both sides of the jaws. Another 19 primary canines had hypoplastic defects, probably the result of unsuccessful extractions. Associated dental abnormalities included hypoplasia of the permanent successors and adjacent primary and permanent teeth, displacement of permanent teeth, midline shift to the extraction side, missing primary lateral incisors (probably accidentally extracted) and distal eruption of permanent lateral incisors, leaving their primary predecessors retained. Parental enquiry revealed that the practice is more common in rural rather than urban areas and still exists in the Ethiopian community in Israel. The findings of this survey should urge the authorities to take steps to stop this practice.

  15. Mosquito larvicidal and biting deterrency activity of bud of Polianthes tuberosa plants extract against Anopheles stephensi and Culex quinquefasciatus.

    Science.gov (United States)

    Anjali, Rawani; Atanu, Banerjee; Goutam, Chandra

    2012-06-01

    Mosquito control by phytochemicals is an alternative method to synthetic insecticides, as it is biodegradable and non resistant to vector mosquito. Polianthes tuberosa is a perennial plant distributed in many parts of India. The present study was undertaken to scientifically evaluate the larvicide and biting deterrency activity of bud of Polianthes tuberosa against Culex quinquefasciatus and Anopheles stephensi. Crude and solvent extract [ethyl acetate, chloroform: methanol (1:1, v/v), acetone] of fresh, mature, bud of P. tuberosa was tested against (ex. quinquefasciatus and An. stephensi. The repellent activity tested by chloroform: methanol (1:1, v/v) solvent extract against both mosquito species. The appropriate lethal concentrations at 24h for chloroform: methanol (1:1, v/v) extract was also studied on non target organisms such as Toxorhynchites larvae, Diplonychus annulatum and Chironomus circumdatus. In a 72 hour bioassay experiment, 0.5% crude extract showed the highest mortality and chloroform: methanol (1:1, v/v) solvent extract showed the highest mortality, the maximum (p larvicide agent. There is no changes in the activity non-target organism so, it is safe to use.

  16. Involvement of Lipid Rafts and Cellular Actin in AcMNPV GP64 Distribution and Virus Budding

    Institute of Scientific and Technical Information of China (English)

    F. J. Haines; C. M. Griffiths; R. D. Possee; C. R. Hawes; L. A. King

    2009-01-01

    GP64 is the major envelope glycoprotein associated with the budded virus (BV) of Autographa californica nucleopolyhedrovirus (AcMNPV) and is essential for attachment and budding of BV particles.Confocal microscopy and flotation assays established the presence of lipid raft domains within the plasma membranes of AcMNPV-infected Sf9 cells and suggested the association of GP64 with lipid rafts during infection. GP64 and filamentous actin (F-actin) were found to co-localise at the cell cortex at 24 and 48 hpi and an additional restructuring of F-actin during infection was visualised, resulting in a strongly polarised distribution of both F-actin and GP64 at the cell cortex. Depletion of F-actin, achieved by treatment of St9 cells with latrunculin B (LB), resulted in the redistribution of GP64 with significant cytoplasmic aggregation and reduced presence at the plasma membrane. Treatment with LB also resulted in reduced production of BV in Sf9 cells. Analysis of virus gene transcription confirmed this reduction was not due to decreased trafficking of nucleocapsids to the nucleus or to decreased production of infectious progeny nucleoeapsids. Reduced BV production due to a lack of GP64 at the plasma membrane of AcMNPV-infected Sf9 cells treated with LB, suggests a key role for F-actin in the egress of BV.

  17. Deep-sequencing transcriptome analysis of field-grown Medicago sativa L. crown buds acclimated to freezing stress.

    Science.gov (United States)

    Song, Lili; Jiang, Lin; Chen, Yue; Shu, Yongjun; Bai, Yan; Guo, Changhong

    2016-09-01

    Medicago sativa L. (alfalfa) 'Zhaodong' is an important forage legume that can safely survive in northern China where winter temperatures reach as low as -30 °C. Survival of alfalfa following freezing stress depends on the amount and revival ability of crown buds. In order to investigate the molecular mechanisms of frost tolerance in alfalfa, we used transcriptome sequencing technology and bioinformatics strategies to analyze crown buds of field-grown alfalfa during winter. We statistically identified a total of 5605 differentially expressed genes (DEGs) involved in freezing stress including 1900 upregulated and 3705 downregulated DEGs. We validated 36 candidate DEGs using qPCR to confirm the accuracy of the RNA-seq data. Unlike other recent studies, this study employed alfalfa plants grown in the natural environment. Our results indicate that not only the CBF orthologs but also membrane proteins, hormone signal transduction pathways, and ubiquitin-mediated proteolysis pathways indicate the presence of a special freezing adaptation mechanism in alfalfa. The antioxidant defense system may rapidly confer freezing tolerance to alfalfa. Importantly, biosynthesis of secondary metabolites and phenylalanine metabolism, which is of potential importance in coordinating freezing tolerance with growth and development, were downregulated in subzero temperatures. The adaptive mechanism for frost tolerance is a complex multigenic process that is not well understood. This systematic analysis provided an in-depth view of stress tolerance mechanisms in alfalfa.

  18. Mateřská škola v Českých Budějovicích

    OpenAIRE

    Šturma, Pavel

    2015-01-01

    Diplomová práce je zpracována ve formě projektové dokumentace k provedení novostavby mateřské školy v Českých Budějovicích. Projektová dokumentace je navržena dle platných norem. Objekt je navržen v katastrálním území České Budějovice. Mateřská škola má dvě nadzemní podlaží. Budova je navržena ze systému Porotherm s kontaktním zateplovacím systémem, zastřešena jednoplášťovou vegetační plochou střechou a založena na základových pasech z prostého betonu. Stropní konstrukce je tvořena z předem p...

  19. Small molecule inhibitors of the Candida albicans budded-to-hyphal transition act through multiple signaling pathways.

    Directory of Open Access Journals (Sweden)

    John Midkiff

    Full Text Available The ability of the pathogenic yeast Candida albicans to interconvert between budded and hyphal growth states, herein termed the budded-to-hyphal transition (BHT, is important for C. albicans development and virulence. The BHT is under the control of multiple cell signaling pathways that respond to external stimuli, including nutrient availability, high temperature, and pH. Previous studies identified 21 small molecules that could inhibit the C. albicans BHT in response to carbon limitation in Spider media. However, the studies herein show that the BHT inhibitors had varying efficacies in other hyphal-inducing media, reflecting their varying abilities to block signaling pathways associated with the different media. Chemical epistasis analyses suggest that most, but not all, of the BHT inhibitors were acting through either the Efg1 or Cph1 signaling pathways. Notably, the BHT inhibitor clozapine, a FDA-approved drug used to treat atypical schizophrenia by inhibiting G-protein-coupled dopamine receptors in the brain, and several of its functional analogs were shown to act at the level of the Gpr1 G-protein-coupled receptor. These studies are the first step in determining the target and mechanism of action of these BHT inhibitors, which may have therapeutic anti-fungal utility in the future.

  20. Genome-wide identification of WRKY family genes in peach and analysis of WRKY expression during bud dormancy.

    Science.gov (United States)

    Chen, Min; Tan, Qiuping; Sun, Mingyue; Li, Dongmei; Fu, Xiling; Chen, Xiude; Xiao, Wei; Li, Ling; Gao, Dongsheng

    2016-06-01

    Bud dormancy in deciduous fruit trees is an important adaptive mechanism for their survival in cold climates. The WRKY genes participate in several developmental and physiological processes, including dormancy. However, the dormancy mechanisms of WRKY genes have not been studied in detail. We conducted a genome-wide analysis and identified 58 WRKY genes in peach. These putative genes were located on all eight chromosomes. In bioinformatics analyses, we compared the sequences of WRKY genes from peach, rice, and Arabidopsis. In a cluster analysis, the gene sequences formed three groups, of which group II was further divided into five subgroups. Gene structure was highly conserved within each group, especially in groups IId and III. Gene expression analyses by qRT-PCR showed that WRKY genes showed different expression patterns in peach buds during dormancy. The mean expression levels of six WRKY genes (Prupe.6G286000, Prupe.1G393000, Prupe.1G114800, Prupe.1G071400, Prupe.2G185100, and Prupe.2G307400) increased during endodormancy and decreased during ecodormancy, indicating that these six WRKY genes may play a role in dormancy in a perennial fruit tree. This information will be useful for selecting fruit trees with desirable dormancy characteristics or for manipulating dormancy in genetic engineering programs. PMID:26951048

  1. Fluorescent Protein-Tagged Sindbis Virus E2 Glycoprotein Allows Single Particle Analysis of Virus Budding from Live Cells.

    Science.gov (United States)

    Jose, Joyce; Tang, Jinghua; Taylor, Aaron B; Baker, Timothy S; Kuhn, Richard J

    2015-12-01

    Sindbis virus (SINV) is an enveloped, mosquito-borne alphavirus. Here we generated and characterized a fluorescent protein-tagged (FP-tagged) SINV and found that the presence of the FP-tag (mCherry) affected glycoprotein transport to the plasma membrane whereas the specific infectivity of the virus was not affected. We examined the virions by transmission electron cryo-microscopy and determined the arrangement of the FP-tag on the surface of the virion. The fluorescent proteins are arranged icosahedrally on the virus surface in a stable manner that did not adversely affect receptor binding or fusion functions of E2 and E1, respectively. The delay in surface expression of the viral glycoproteins, as demonstrated by flow cytometry analysis, contributed to a 10-fold reduction in mCherry-E2 virus titer. There is a 1:1 ratio of mCherry to E2 incorporated into the virion, which leads to a strong fluorescence signal and thus facilitates single-particle tracking experiments. We used the FP-tagged virus for high-resolution live-cell imaging to study the spatial and temporal aspects of alphavirus assembly and budding from mammalian cells. These processes were further analyzed by thin section microscopy. The results demonstrate that SINV buds from the plasma membrane of infected cells and is dispersed into the surrounding media or spread to neighboring cells facilitated by its close association with filopodial extensions.

  2. Deep-sequencing transcriptome analysis of field-grown Medicago sativa L. crown buds acclimated to freezing stress.

    Science.gov (United States)

    Song, Lili; Jiang, Lin; Chen, Yue; Shu, Yongjun; Bai, Yan; Guo, Changhong

    2016-09-01

    Medicago sativa L. (alfalfa) 'Zhaodong' is an important forage legume that can safely survive in northern China where winter temperatures reach as low as -30 °C. Survival of alfalfa following freezing stress depends on the amount and revival ability of crown buds. In order to investigate the molecular mechanisms of frost tolerance in alfalfa, we used transcriptome sequencing technology and bioinformatics strategies to analyze crown buds of field-grown alfalfa during winter. We statistically identified a total of 5605 differentially expressed genes (DEGs) involved in freezing stress including 1900 upregulated and 3705 downregulated DEGs. We validated 36 candidate DEGs using qPCR to confirm the accuracy of the RNA-seq data. Unlike other recent studies, this study employed alfalfa plants grown in the natural environment. Our results indicate that not only the CBF orthologs but also membrane proteins, hormone signal transduction pathways, and ubiquitin-mediated proteolysis pathways indicate the presence of a special freezing adaptation mechanism in alfalfa. The antioxidant defense system may rapidly confer freezing tolerance to alfalfa. Importantly, biosynthesis of secondary metabolites and phenylalanine metabolism, which is of potential importance in coordinating freezing tolerance with growth and development, were downregulated in subzero temperatures. The adaptive mechanism for frost tolerance is a complex multigenic process that is not well understood. This systematic analysis provided an in-depth view of stress tolerance mechanisms in alfalfa. PMID:27272950

  3. Fluorescent Protein-Tagged Sindbis Virus E2 Glycoprotein Allows Single Particle Analysis of Virus Budding from Live Cells

    Directory of Open Access Journals (Sweden)

    Joyce Jose

    2015-11-01

    Full Text Available Sindbis virus (SINV is an enveloped, mosquito-borne alphavirus. Here we generated and characterized a fluorescent protein-tagged (FP-tagged SINV and found that the presence of the FP-tag (mCherry affected glycoprotein transport to the plasma membrane whereas the specific infectivity of the virus was not affected. We examined the virions by transmission electron cryo-microscopy and determined the arrangement of the FP-tag on the surface of the virion. The fluorescent proteins are arranged icosahedrally on the virus surface in a stable manner that did not adversely affect receptor binding or fusion functions of E2 and E1, respectively. The delay in surface expression of the viral glycoproteins, as demonstrated by flow cytometry analysis, contributed to a 10-fold reduction in mCherry-E2 virus titer. There is a 1:1 ratio of mCherry to E2 incorporated into the virion, which leads to a strong fluorescence signal and thus facilitates single-particle tracking experiments. We used the FP-tagged virus for high-resolution live-cell imaging to study the spatial and temporal aspects of alphavirus assembly and budding from mammalian cells. These processes were further analyzed by thin section microscopy. The results demonstrate that SINV buds from the plasma membrane of infected cells and is dispersed into the surrounding media or spread to neighboring cells facilitated by its close association with filopodial extensions.

  4. The effects of heat treatment on physical properties and surface roughness of red-bud maple (Acer trautvetteri Medw.) wood.

    Science.gov (United States)

    Korkut, Derya Sevim; Guller, Bilgin

    2008-05-01

    Heat treatment is often used to improve the dimensional stability of wood. In this study, the effects of heat treatment on physical properties and surface roughness of red-bud maple (Acer trautvetteri Medw.) wood were examined. Samples obtained from Düzce Forest Enterprises, Turkey, were subjected to heat treatment at varying temperatures and durations. The physical properties of heat-treated samples were compared against controls in order to determine their; oven-dry density, air-dry density, and swelling properties. A stylus method was employed to evaluate the surface characteristics of the samples. Roughness measurements, using the stylus method, were made in the direction perpendicular to the fiber. Three main roughness parameters; mean arithmetic deviation of profile (Ra), mean peak-to-valley height (Rz), and maximum roughness (Rmax) obtained from the surface of wood, were used to evaluate the effect of heat treatment on the surface characteristics of the specimens. Significant differences were determined (p>0.05) between surface roughness parameters (Ra, Rz, Rmax) at three different temperatures and three periods of heat treatment. The results showed that the values of density, swelling and surface roughness decreased with increasing temperature treatment and treatment times. Red-bud maple wood could be utilized successfully by applying proper heat treatment techniques without any losses in investigated parameters. This is vital in areas, such as window frames, where working stability and surface smoothness are important factors.

  5. Headspace Hanging Drop Liquid Phase Microextraction and Gas Chromatography-Mass Spectrometry for the Analysis of Flavors from Clove Buds

    International Nuclear Information System (INIS)

    A novel sample pretreatment technique, headspace hanging drop liquid phase microextraction (HS-LPME) was studied and applied to the determination of flavors from solid clove buds by gas chromatography-mass spectrometry (GC-MS). Several parameters affecting on HS-LPME such as organic solvent drop volume, extraction time, extraction temperature and phase ratio were investigated. 1-Octanol was selected as the extracting solvent, drop size was fixed to 0.6 μL. 60 min extraction time at 25 .deg. C was chosen. HS-LPME has the good efficiency demonstrated by the higher partition equilibrium constant (Klh) values and concentration factor (CF) values. The limits of detection (LOD) were 1.5-3.2 ng. The amounts of eugenol, β-caryophyllene and eugenol acetate from the clove bud sample were 1.90 mg/g, 1.47 mg/g and 7.0 mg/g, respectively. This hanging drop based method is a simple, fast and easy sample enrichment technique using minimal solvent. HSLPME is an alternative sample preparation method for the analysis of volatile aroma compounds by GC-MS

  6. Evidence against mediation of adenosine-3',5'-cyclic monophosphate in the bud-inducing effect of cytokinins in moss protonemata

    Directory of Open Access Journals (Sweden)

    J. Scheneider

    2015-05-01

    Full Text Available Effects Oif adenosdne-3',5'-cyclic monophosphate (cAMP, N6,O2-dibuityryl adenosine-3',5'-cyclic monophosphate (DBcAMP, caffeine and theophylline on the bud-inducing activity of cytokinin in the protonema of two moss species, Ceratodon purpureus and Funaria hygrometrica were examined. The sub-stances have been found ineffective as gametophore bud inducers. Some synergism between cytokinin and cAMP or DBcAMP was observed with relation to the buds' growth, but this effect is nonspecific since it can be obtained with 5'-AMP or 5'-GMiP as well, The results seem to exclude the possibility of an involvement of cAMP as a second messenger in the mechanism of cytokinin action on morphogenetic processes in moss protonemata.

  7. A density functional reactivity theory (DFRT) based approach to understand the effect of symmetry of fullerenes on the kinetic, thermodynamic and structural aspects of carbon NanoBuds

    Science.gov (United States)

    Sarmah, Amrit; Roy, Ram Kinkar

    2016-06-01

    In the present study, we have rationalized the effect of variation in the symmetry of relatively smaller fullerene (C32) on the mode of its interaction with semi-conducting Single-Walled Carbon Nanotubes (SWCNTs) in the process of formation of stable hybrid carbon NanoBuds. Thermodynamic and kinetic parameters, along with the charge transfer values associated with the interaction between fullerene and SWCNTs, have been evaluated using an un-conventional and computationally cost-effective method based on density functional reactivity theory (DFRT). In addition to this, conventional DFT based studies are also performed to substantiate the growth of NanoBud structures formed by the interaction between fullerene and SWCNTs. The findings of the present study suggest that the kinetic, thermodynamic and structural aspects of hybrid carbon NanoBuds are significantly influenced by both the symmetry of C32 fullerene and its site of covalent attachment to the SWCNT.

  8. Fumigant activity of plant essential oils and components from garlic (Allium sativum) and clove bud (Eugenia caryophyllata) oils against the Japanese termite (Reticulitermes speratus Kolbe).

    Science.gov (United States)

    Park, Il-Kwon; Shin, Sang-Chul

    2005-06-01

    Plant essential oils from 29 plant species were tested for their insecticidal activities against the Japanese termite, Reticulitermes speratus Kolbe, using a fumigation bioassay. Responses varied with plant material, exposure time, and concentration. Good insecticidal activity against the Japanese termite was achived with essential oils of Melaleuca dissitiflora, Melaleuca uncinata, Eucalyptus citriodora, Eucalyptus polybractea, Eucalyptus radiata, Eucalyptus dives, Eucalyptus globulus, Orixa japonica, Cinnamomum cassia, Allium cepa, Illicium verum, Evodia officinalis, Schizonepeta tenuifolia, Cacalia roborowskii, Juniperus chinensis var. horizontalis, Juniperus chinensis var. kaizuka, clove bud, and garlic applied at 7.6 microL/L of air. Over 90% mortality after 3 days was achieved with O. japonica essential oil at 3.5 microL/L of air. E. citriodora, C. cassia, A. cepa, I. verum, S. tenuifolia, C. roborowskii, clove bud, and garlic oils at 3.5 microL/L of air were highly toxic 1 day after treatment. At 2.0 microL/L of air concentration, essential oils of I. verum, C. roborowskik, S. tenuifolia, A. cepa, clove bud, and garlic gave 100% mortality within 2 days of treatment. Clove bud and garlic oils showed the most potent antitermitic activity among the plant essential oils. Garlic and clove bud oils produced 100% mortality at 0.5 microL/L of air, but this decreased to 42 and 67% after 3 days of treatment at 0.25 microL/L of air, respectively. Analysis by gas chromatography-mass spectrometry led to the identification of three major compounds from garlic oil and two from clove bud oils. These five compounds from two essential oils were tested individually for their insecticidal activities against Japanese termites. Responses varied with compound and dose. Diallyl trisulfide was the most toxic, followed by diallyl disulfide, eugenol, diallyl sulfide, and beta-caryophyllene. The essential oils described herein merit further study as potential fumigants for termite

  9. Live cell imaging of the assembly, disassembly, and actin cable–dependent movement of endosomes and actin patches in the budding yeast, Saccharomyces cerevisiae

    OpenAIRE

    Huckaba, Thomas M.; Gay, Anna Card; Pantalena, Luiz Fernando; Yang, Hyeong-Cheol; Liza A Pon

    2004-01-01

    Using FM4-64 to label endosomes and Abp1p-GFP or Sac6p-GFP to label actin patches, we find that (1) endosomes colocalize with actin patches as they assemble at the bud cortex; (2) endosomes colocalize with actin patches as they undergo linear, retrograde movement from buds toward mother cells; and (3) actin patches interact with and disassemble at FM4-64–labeled internal compartments. We also show that retrograde flow of actin cables mediates retrograde actin patch movement. An Arp2/3 complex...

  10. Srovnání MHD v Českých Budějovicích a v Hradci Králové

    OpenAIRE

    Fialová, Zdeňka

    2008-01-01

    Charakteristika městské hromadné dopravy a základní pojmy;města České Budějovice a Hradec Králové;historie MHD;současné provozování MHD v obou městech;dopravní síť;dopravní dostupnost;přepravní tarif;celkové srovnání MHD v Českých Budějovicích a v Hradci Králové

  11. Crosstalk among Taenia crassiceps (ORF Strain) cysts regulates their rates of budding by ways of soluble and contact signals exchanged between them.

    Science.gov (United States)

    Marcela, Esquivel-Velázquez; Ricardo, Hernández; Carlos, Larralde; Pedro, Ostoa-Saloma

    2014-01-01

    Herein we report that in vitro experiments with different initial parasite densities (1, 5, and 10 cysts per mL of culture medium) show that cysts at densities of 10 and 5 grow faster than those at 1, and that they release into the culture medium factors which increase the budding rates of the slower lower-density ones. Close contact among the incubated cysts also favors budding, thus suggesting the participation of surface sensors of parasite crowding. Thus, contact signals, together with the release of soluble growth factors, could endow cysts with the capacity to sense and regulate their numbers inside their habitat in relation to their population density.

  12. Žena v Českých Budějovicích ve světle testamentů

    OpenAIRE

    MRVKOVÁ, Lucie

    2014-01-01

    The Bachelor thesis "Women of town České Budějovice in the light of testaments" proceses the last wills and the give up the possesion from 14931550, which are deposited in SOkA České Budějovice. Another used source is the town lawcode from Brikcí of Licko, which is editorially processed. The attention is devoted to inheritance and property law of townwomen in default standard. From testaments is made a picture of women in role of women-testators, heiress and trusteers. The thesis includes the...

  13. Stable Pseudohyphal Growth in Budding Yeast Induced by Synergism between Septin Defects and Altered MAP-kinase Signaling.

    Science.gov (United States)

    Kim, Junwon; Rose, Mark D

    2015-12-01

    Upon nutrient limitation, budding yeasts like Saccharomyces cerevisiae can be induced to adopt alternate filament-like growth patterns called diploid pseudohyphal or invasive haploid growth. Here, we report a novel constitutive pseudohyphal growth state, sharing some characteristics with classic forms of filamentous growth, but differing in crucial aspects of morphology, growth conditions and genetic regulation. The constitutive pseudohyphal state is observed in fus3 mutants containing various septin assembly defects, which we refer to as sadF growth (septin assembly defect induced filamentation) to distinguish it from classic filamentation pathways. Similar to other filamentous states, sadF cultures comprise aggregated chains of highly elongated cells. Unlike the classic pathways, sadF growth occurs in liquid rich media, requiring neither starvation nor the key pseudohyphal proteins, Flo8p and Flo11p. Moreover sadF growth occurs in haploid strains of S288C genetic background, which normally cannot undergo pseudohyphal growth. The sadF cells undergo highly polarized bud growth during prolonged G2 delays dependent on Swe1p. They contain septin structures distinct from classical pseudo-hyphae and FM4-64 labeling at actively growing tips similar to the Spitzenkörper observed in true hyphal growth. The sadF growth state is induced by synergism between Kss1p-dependent signaling and septin assembly defects; mild disruption of mitotic septins activates Kss1p-dependent gene expression, which exacerbates the septin defects, leading to hyper-activation of Kss1p. Unlike classical pseudo-hyphal growth, sadF signaling requires Ste5, Ste4 and Ste18, the scaffold protein and G-protein β and γ subunits from the pheromone response pathway, respectively. A swe1 mutation largely abolished signaling, breaking the positive feedback that leads to amplification of sadF signaling. Taken together, our findings show that budding yeast can access a stable constitutive pseudohyphal growth

  14. Stable Pseudohyphal Growth in Budding Yeast Induced by Synergism between Septin Defects and Altered MAP-kinase Signaling.

    Directory of Open Access Journals (Sweden)

    Junwon Kim

    2015-12-01

    Full Text Available Upon nutrient limitation, budding yeasts like Saccharomyces cerevisiae can be induced to adopt alternate filament-like growth patterns called diploid pseudohyphal or invasive haploid growth. Here, we report a novel constitutive pseudohyphal growth state, sharing some characteristics with classic forms of filamentous growth, but differing in crucial aspects of morphology, growth conditions and genetic regulation. The constitutive pseudohyphal state is observed in fus3 mutants containing various septin assembly defects, which we refer to as sadF growth (septin assembly defect induced filamentation to distinguish it from classic filamentation pathways. Similar to other filamentous states, sadF cultures comprise aggregated chains of highly elongated cells. Unlike the classic pathways, sadF growth occurs in liquid rich media, requiring neither starvation nor the key pseudohyphal proteins, Flo8p and Flo11p. Moreover sadF growth occurs in haploid strains of S288C genetic background, which normally cannot undergo pseudohyphal growth. The sadF cells undergo highly polarized bud growth during prolonged G2 delays dependent on Swe1p. They contain septin structures distinct from classical pseudo-hyphae and FM4-64 labeling at actively growing tips similar to the Spitzenkörper observed in true hyphal growth. The sadF growth state is induced by synergism between Kss1p-dependent signaling and septin assembly defects; mild disruption of mitotic septins activates Kss1p-dependent gene expression, which exacerbates the septin defects, leading to hyper-activation of Kss1p. Unlike classical pseudo-hyphal growth, sadF signaling requires Ste5, Ste4 and Ste18, the scaffold protein and G-protein β and γ subunits from the pheromone response pathway, respectively. A swe1 mutation largely abolished signaling, breaking the positive feedback that leads to amplification of sadF signaling. Taken together, our findings show that budding yeast can access a stable constitutive

  15. Nematodes from galls on Myrtaceae. VIII. Fergusobia from small galls on shoot buds, with descriptions of four new species.

    Science.gov (United States)

    Davies, Kerrie A; Bartholomaeus, Faerlie; Giblin-Davis, Robin M; Ye, Weimin; Taylor, Gary S; Thomas, W Kelley

    2014-08-28

    Small shoot bud galls induced by the Fergusobia (Nematoda: Neotylenchidae)/Fergusonina (Diptera: Fergusoninidae) mutualism occur on various Eucalyptus spp. Four new species of Fergusobia, collected from small shoot bud galls on Eucalyptus camaldulensis, E. gomphocephala and E. leucoxylon, are described. Fergusobia gomphocephalae Davies n. sp. is morphologically characterized by a combination of a small C-shaped parthenogenetic female with a variable, conoid tail, a small C-shaped infective female with a hemispherical tail tip, and an arcuate or J-shaped male with a broad tail, angular spicule and short peloderan bursa. Fergusobia leucoxylonae Davies n. sp. has a C-shaped parthenogenetic female with a conoid tail with a narrowly rounded tip, an arcuate infective female with a broadly rounded tail tip, and an almost straight to barely J-shaped male with angular (not heavily sclerotised) spicule and short bursa. Fergusobia schmidti Davies & Bartholomaeus n. sp. has an arcuate to open C-shaped parthenogenetic female with a relatively large body diameter, relatively long stylet and small tail with a broadly rounded tail tip, an open C-shaped infective female with a broadly rounded to hemispherical tail tip, and an arcuate to barely J-shaped male with spicules angular at about 33% of their length and peloderan bursa arising at about half body length. Fergusobia sporangae Davies n. sp. has an arcuate to open C-shaped parthenogenetic female with a relatively long stylet and a broadly rounded tail tip, an arcuate infective female with a short tail with a broadly rounded to hemispherical tip, and an arcuate to barely J-shaped male with angular (not heavily sclerotised) spicule and short peloderan bursa. Various forms of small shoot bud galls are described. From phylogenetic analyses based on sequences of the D2/D3 expansion segment of the large subunit rRNA gene, the four new species belong to two sister clades of Fergusobia. The larval shield morphology of their associated

  16. Effects of carbon ion beam irradiation on the shoot regeneration from in vitro axillary bud explants of the Impatiens hawkeri

    Science.gov (United States)

    Zhou, Libin; Zhou, Libin; Li, Wenjian; Li, Ping; Dong, Xicun; Qu, Ying; Ma, Shuang; Li, Qiang

    Accelerated ion beams is an excellent mutagen in plant breeding which can induce higher mutation frequencies and wider mutation spectrum than those of low linear energy transfer (LET) irradiations, such as X-rays (Okamura et al. 2003, Yamaguchi et al. 2003). Mutation breeding operation of two Saintpaulia ionahta cultivars using the method combining plant tissue culture technique and carbon ion beam irradiations were set out at Institute of Modern Physics from 2005 (Zhou et al. 2006). The effects of 960 MeV carbon ion beam and 8 MeV X-ray irradiations on regenerated shoots of Impatiens hawkeri from another kind of explants named in vitro axillary buds explants were studied recently. The biology endpoints in this study included relative number of roots (RNR), relative length of roots (RLR), relative height of shoots (RHS), relative number of nodes (RNN), survival fraction (SF) and morphology changes in the regenerated shoots. The experimental results showed that carbon ion beams inhibited the root and stem developments of axillary bud explants more severely than X-rays did. And the 50% lethal dose (LD50 ) is about 23.3 Gy for the carbon ion beam and 49.1 Gy for the X-rays, respectively. Relative biological effectiveness (RBE) of Impatiens hawkeri with respect to X-rays according to 50% SF was about two. Secondly, the percentage of shoots regenerated with malformed shoots including curliness, carnification, nicks in all Impatiens hawkeri axillary bud explants irradiated with carbon ion beam at 20 Gy accounted for 55.6%, while the highest number for the 40 Gy X-ray irradiation was 40%. Last, many regenerated shoots whose vascular bundle fused together were obtained only from explants irradiated with carbon ion beams. Based on the results above, it can be concluded that the effect of mutation induction by carbon ion beam irradiation on the axillary explants of Impatiens hawkeri is better than that by X-ray irradiation; and the optimal mutagenic dose varies from 20 Gy

  17. Tracheid production phenology of Picea mariana and its relationship with climatic fluctuations and bud development using multivariate analysis.

    Science.gov (United States)

    Dufour, Boris; Morin, Hubert

    2010-07-01

    Research on cambium phenology in trees and its limiting factors in natural conditions is still at an early stage of development, restricting our capacity to precisely evaluate the effect of growing season length and climate fluctuations on tracheid production. The first objective of this paper was to describe cambial tracheid production phenology of black spruce (Picea mariana (Mills.) BSP). Repeated tree ring sampling was performed from 2002 to 2006 on four sites (48 degrees 13.78' N, 71 degrees 15.18' W; 48 degrees 51.92' N, 70 degrees 20.57' W; 49 degrees 43.92' N, 71 degrees 56.88' W; and 50 degrees 41.78' N, 72 degrees 11.03' W) representative of closed black spruce forest in Quebec, Canada. The timing of cambial initiation and cambial cessation in black spruce differs from year to year, the first occurring on 4 June on average, whereas the second occurs on 15 August. During a single year, these events do not vary significantly in space within the study area. The duration of cambial tracheid production does not vary significantly in either time or space. The second objective of this study was to identify the climatic factors that explain variations in initiation and cessation. Air temperature and humidity, soil temperature and water content, rain precipitations, snow cover as well as photosynthetically active radiation were monitored at each studied site. These were then used to create sets of candidate regressors to explain timing of phenological events. Timing of cambial initiation is primarily dependent on mean temperature between mid-March and initiation itself. Vapor pressure during this period is also important but in a negative way. A significant effect of the previous year's August soil and air temperature conditions suggests a link with spring bud activity resumption, an interpretation that is supported by an analysis significantly linking measured timing of bud break to cambial initiation. Cessation of cambial tracheid production is influenced by

  18. Tracing the allocation of recently assimilated C into key metabolites in Norway spruce (Picea abies) shortly after bud break

    Science.gov (United States)

    Heinrich, Steffen; Dippold, Michaela; Werner, Christiane; Wiesenberg, Guido; Kuzyakov, Yakov; Glaser, Bruno

    2015-04-01

    Plants allocate carbon (C) to sink tissues depending on phenological, physiological or environmental factors. We still have little knowledge on C partitioning into various cellular compounds and metabolic pathways, especially during tree growth after bud break. Here we investigated C partitioning of freshly assimilated C in Norway spruce by in-situ 13C short-term pulse labeling 15 days after bud break. We quantified 13C incorporation into tree compartments (needles, branches, stem) and into water soluble organic carbon (WSOC) by elemental analyzer-isotope ratio mass spectrometry (EA-IRMS). In addition, we determined 13C allocation into key metabolites of amino acids, hemicellulose sugars, fatty acids and alkanes by compound-specific 13C analysis via gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). The 13C allocation within the trees reflected the needles as major C sink accounting for 86% of the freshly assimilated C. After 6 h 13C was distributed over a broad spectrum of plant metabolites but not homogenously. Highest allocation was observed into structurally relevant compound classes of hemicellulose-derived sugars and proteinogenic amino acids (0.6% and 10% of needle 13C, respectively). However, needle growth also caused high C allocation into pathways not involved in formation of structural compounds like pathways in secondary metabolism, C-1 metabolism or amino acid synthesis from photorespiratory acitivity. C allocation into such pathways could be identified due to the high enrichment of key metabolites within the amino acids. In addition, high 13C allocation was found into the n-alkyl lipid biosynthesis (0.2 % of needle 13C) with 1) higher allocation into intracellular than cuticular fatty acids, presumably for thylakoide membrane formation and 2) decreasing 13C allocation along the lipid transformation and translocation pathways (precursor fatty acids (C16 & C18) > elongated long chain fatty acids > decarbonylated n

  19. Phytochemical, Physico-chemical & Spectroscopic Characteristics of Ethanolic Extract of Leaf, Stem and Flower bud of Hibiscus hispidissimus Griffith

    Directory of Open Access Journals (Sweden)

    Thamizh Selvam N

    2015-08-01

    Full Text Available The plant Hibiscus hispidissimus belongs to the family Malvaceae (Mallow family. The plant has wide range of medicinal uses. Considering the ethno medicinal value of Hibiscus hispidissimus, the present work has been taken up to document the physico-chemical composition, phytochemical details and spectrophotometric characteristics of the plant. The work has been carried out on ethanolic extract of leaf, stem and flower bud of H. hispidissimus. Phytochemical analysis showed the presence of saponis, tannins, glycosides, diterpenes and quinones. Spectroscopic characteristics were analyzed and found to have wide range of compounds including steroids, alkaloids, pigments like chlorophyll a and b, phenolic compounds mainly gallic acid, flavanoids like anthocyanins, flavanols, flvanones and isoflavones.

  20. Effects of soil water deficits on three genotypes of potted Campanula medium plants during bud formation stage

    DEFF Research Database (Denmark)

    Mao, Hongyu; Sun, Yanqi; Müller, Renate;

    2014-01-01

    of T in G100 would allow the plant to conserve water in the soil; however, owing to its putative larger leaf area and greater stomatal density on the adaxial leaf surface, G100 quickly depleted soil water and resulted in severe drought stress to the plants as exemplified by the significantly lowered gs......Potted ornamental plants are often exposed to drought stress during shipping and retailing, which decreases the value and postharvest quality. Thus, selection of genotypes which can better withstand soil water deficits is essential for sustainable production. Here, the response of three genotypes...... of potted Campanula medium (denoted as G100, G102 and G104) to progressive soil drying was investigated and their post-production performance was evaluated for four weeks. The potted plants were grown in a climate controlled greenhouse and were either well-watered (W) or drought-stressed (D) at floral bud...

  1. Transformation of Chinese Cabbage (Brassica rapa L. ssp.pekinensis) by Agrobacterium Micro-Injection into Flower Bud

    Institute of Scientific and Technical Information of China (English)

    YAN Ji-yong; HE Yu-ke; CAO Jia-shu

    2003-01-01

    We obtained two lines of Chinese head cabbage (Brassica rapa L. ssp. pekinensis) selfed progenies containing both an anti-sense gene of BcpLH and a gene for resistance to kanamycin by micro-injecting buds of their primary transformants (T0) with Agrobacterium tumefaciens strain LBA4404. 31 positive plants resistant to kanamycien were recovered. Southern blot analysis confirmed the presence of T-DNA in two transgenic plants. One (DHZ-13-1) exhibits the characteristics of out-toward rosette and cauline leaves, and nested flower model in which secondary complete flower developed from the base of the primary ovary and the third flower from the ovary in the secondary flower, and so on, while another(DHZ-6-1) has no phenotype change. ABA and IAA affected the root growth of progeny of DHZ-13-1, but 6-BA was insensitive to hypocotyl growth during its seedling development.

  2. A Geometrically-Constrained Mathematical Model of Mammary Gland Ductal Elongation Reveals Novel Cellular Dynamics within the Terminal End Bud.

    Directory of Open Access Journals (Sweden)

    Ingrid Paine

    2016-04-01

    Full Text Available Mathematics is often used to model biological systems. In mammary gland development, mathematical modeling has been limited to acinar and branching morphogenesis and breast cancer, without reference to normal duct formation. We present a model of ductal elongation that exploits the geometrically-constrained shape of the terminal end bud (TEB, the growing tip of the duct, and incorporates morphometrics, region-specific proliferation and apoptosis rates. Iterative model refinement and behavior analysis, compared with biological data, indicated that the traditional metric of nipple to the ductal front distance, or percent fat pad filled to evaluate ductal elongation rate can be misleading, as it disregards branching events that can reduce its magnitude. Further, model driven investigations of the fates of specific TEB cell types confirmed migration of cap cells into the body cell layer, but showed their subsequent preferential elimination by apoptosis, thus minimizing their contribution to the luminal lineage and the mature duct.

  3. Antioxidant and antimicrobial activities of clove bud essential oil and eugenol nanoparticles in alcohol-free microemulsion.

    Science.gov (United States)

    Hamed, Said Fatouh; Sadek, Zainab; Edris, Amr

    2012-01-01

    Clove bud essential oil (CEO) and its major individual phenolic constituent eugenol were formulated as nanoparticles in water-based microemulsion systems. The oil titration method was used to incorporate different amounts of the oil and eugenol in the micellar solution of Tween-20. The Antioxidant and antimicrobial activities were evaluated using the DPPH* free radical scavenging assay and the agar disc dilution method, respectively. Results showed that microemulsion improved the evaluated activities of CEO and eugenol compared with the crude counterparts. Individual eugenol microemulsion was more effective than CEO microemulsion which contained only 61.7% eugenol among its constituents. The results of this study could have potential applications in water-based disinfectants, preservation and flavoring of food and in personal hygiene products. It may also have promising applications in the nutraceutical and functional beverage field. PMID:23138253

  4. Molecular profiling of tumour budding implicates TGFβ-mediated epithelial–mesenchymal transition as a therapeutic target in oral squamous cell carcinoma

    DEFF Research Database (Denmark)

    Jensen, David Hebbelstrup; Dabelsteen, Erik; Specht, Lena;

    2015-01-01

    collected from oral squamous cell carcinoma (OSCC) specimens using laser capture microdissection and examined with RNA sequencing and miRNA-qPCR arrays. Compared with cells from the central parts of the tumours, budding cells exhibited a particular gene expression signature comprising factors involved...

  5. ESCRT-III CHMP2A and CHMP3 form variable helical polymers in vitro and act synergistically during HIV-1 budding.

    Science.gov (United States)

    Effantin, Grégory; Dordor, Aurélien; Sandrin, Virginie; Martinelli, Nicolas; Sundquist, Wesley I; Schoehn, Guy; Weissenhorn, Winfried

    2013-02-01

    The endosomal sorting complex required for transport-III (ESCRT-III) proteins are essential for budding of some enveloped viruses, for the formation of intraluminal vesicles at the endosome and for the abscission step of cytokinesis. ESCRT-III proteins form polymers that constrict membrane tubes, leading to fission. We have used electron cryomicroscopy to determine the molecular organization of pleiomorphic ESCRT-III CHMP2A-CHMP3 polymers. The three-dimensional reconstruction at 22 Å resolution reveals a helical organization of filaments of CHMP molecules organized in a head-to-tail fashion. Protease susceptibility experiments indicate that polymerization is achieved via conformational changes that increase the protomer stability. Combinatorial siRNA knockdown experiments indicate that CHMP3 contributes synergistically to HIV-1 budding, and the CHMP3 contribution is ~ 10-fold more pronounced in concert with CHMP2A than with CHMP2B. This is consistent with surface plasmon resonance affinity measurements that suggest sequential CHMP4B-CHMP3-CHMP2A recruitment while showing that both CHMP2A and CHMP2B interact with CHMP4B, in agreement with their redundant functions in HIV-1 budding. Our data thus indicate that the CHMP2A-CHMP3 polymer observed in vitro contributes to HIV-1 budding by assembling on CHMP4B polymers. PMID:23051622

  6. Alteration of starch hydrolyzing enzyme inhibitory properties, antioxidant activities, and phenolic profile of clove buds (Syzygium aromaticum L.) by cooking duration.

    Science.gov (United States)

    Adefegha, Stephen A; Oboh, Ganiyu; Oyeleye, Sunday I; Osunmo, Kolawole

    2016-03-01

    This study assessed the effect of cooking duration on starch hydrolyzing enzyme (α-amylase and α-glucosidase) activities, antioxidant (1,1-diphenyl-2 picrylhydrazyl [DPPH*], hydroxyl [OH*] radicals scavenging abilities and reducing power) properties, and phenolic profile of clove buds. Clove buds (raw) were cooked for 10 (SC 10) and 20 min (SC 20) and subsequently, their effects were assessed on enzyme activities, antioxidant properties, and phenolic profile. Inhibition of α-amylase and α-glucosidase activities and radicals scavenging abilities were altered by cooking in the trend; raw SC 20, with IC 50 values ranging from 0.25 to 0.52 mg/mL and 0.10 to 1.50 mg/mL respectively. HPLC phenolic profile of the clove buds revealed significant (P < 0.05) changes in the amount of chlorogenic acid, quercitrin, quercetin, and kaempferol at different cooking duration. Thus, cooking duration may alter the phenolic compositions and nutraceutical potentials of clove bud by activation and/or deactivation of redox-active metabolites. PMID:27004114

  7. Primary culture and morphologic observation on the taste bud cells in mice%小鼠味蕾细胞形态观察与原代培养

    Institute of Scientific and Technical Information of China (English)

    胡云霞; 柴琳

    2011-01-01

    目的:分离、培养小鼠味蕾细胞.方法:以小白鼠为研究对象,取其舌部组织,制成石蜡切片,从舌上皮分离出味蕾细胞置于Ⅰ型鼠尾胶原包被的含复合培养基的培养板上进行原代培养.结果:显微镜下培养的味蕾细胞与石蜡切片中味蕾细胞形态相似.结论:通过酶消化法可以进行小鼠味蕾细胞原代培养,并为后续味蕾细胞及苦味物质研究奠定技术基础.%Objective: To isolate and culture mouse taste bud cells.Methods: The tongue tissues were obtained from the mice by preparing the paraffin serial sections to isolate the taste bud cells from the lingual epithelia, and the cells were cultured in the plates coated with rat tail tendon collagen type Ⅰ complex medium for primary culture.Results: Microscopically, the taste bud cells resembled the cells from paraffin sections.Conclusion: The taste bud cells of mouse may be primarily cultured by enzyme digestion, which potentially lays the foundation for further study of the cells and taste perception of bitter element.

  8. Lrp4 regulates initiation of ureteric budding and is crucial for kidney formation--a mouse model for Cenani-Lenz syndrome.

    Directory of Open Access Journals (Sweden)

    Courtney M Karner

    Full Text Available BACKGROUND: Development of the kidney is initiated when the ureteric bud (UB branches from the Wolffian duct and invades the overlying metanephric mesenchyme (MM triggering the mesenchymal/epithelial interactions that are the basis of organ formation. Multiple signaling pathways must be integrated to ensure proper timing and location of the ureteric bud formation. METHODS AND PRINCIPAL FINDINGS: We have used gene targeting to create an Lrp4 null mouse line. The mutation results in early embryonic lethality with a subpenetrant phenotype of kidney agenesis. Ureteric budding is delayed with a failure to stimulate the metanephric mesenchyme in a timely manner, resulting in failure of cellular differentiation and resulting absence of kidney formation in the mouse as well as comparable malformations in humans with Cenani-Lenz syndrome. CONCLUSION: Lrp4 is a multi-functional receptor implicated in the regulation of several molecular pathways, including Wnt and Bmp signaling. Lrp4(-/- mice show a delay in ureteric bud formation that results in unilateral or bilateral kidney agenesis. These data indicate that Lrp4 is a critical regulator of UB branching and lack of Lrp4 results in congenital kidney malformations in humans and mice.

  9. ALTERED SENSITIVITY OF THE MOUSE FETUS TO IMPAIRED PROSTATIC BUD FORMATION BY DIOXIN: INFLUENCE OF GENETIC BACKGROUND AND NULL EXPRESSION OF TGF-ALFA AND EGF

    Science.gov (United States)

    Altered sensitivity of the mouse fetus to impaired prostatic bud formation by dioxin: Influence of genetic background and null expression of TGF and EGF. Rasmussen, N.T., Lin T-M., Fenton, S.E., Abbott, B.D. and R.E. Peterson. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)...

  10. The putative Notch ligand HyJagged is a transmembrane protein present in all cell types of adult Hydra and upregulated at the boundary between bud and parent

    Directory of Open Access Journals (Sweden)

    Tischer Susanne

    2011-09-01

    Full Text Available Abstract Background The Notch signalling pathway is conserved in pre-bilaterian animals. In the Cnidarian Hydra it is involved in interstitial stem cell differentiation and in boundary formation during budding. Experimental evidence suggests that in Hydra Notch is activated by presenilin through proteolytic cleavage at the S3 site as in all animals. However, the endogenous ligand for HvNotch has not been described yet. Results We have cloned a cDNA from Hydra, which encodes a bona-fide Notch ligand with a conserved domain structure similar to that of Jagged-like Notch ligands from other animals. Hyjagged mRNA is undetectable in adult Hydra by in situ hybridisation but is strongly upregulated and easily visible at the border between bud and parent shortly before bud detachment. In contrast, HyJagged protein is found in all cell types of an adult hydra, where it localises to membranes and endosomes. Co-localisation experiments showed that it is present in the same cells as HvNotch, however not always in the same membrane structures. Conclusions The putative Notch ligand HyJagged is conserved in Cnidarians. Together with HvNotch it may be involved in the formation of the parent-bud boundary in Hydra. Moreover, protein distribution of both, HvNotch receptor and HyJagged indicate a more widespread function for these two transmembrane proteins in the adult hydra, which may be regulated by additional factors, possibly involving endocytic pathways.

  11. Molecular detection assay of the bud mite Trisetacus juniperinus on Cupressus sempervirens in nurseries of central Italy.

    Science.gov (United States)

    Bouneb, Mabrouk; de Lillo, Enrico; Roversi, Pio Federico; Simoni, Sauro

    2014-02-01

    Trisetacus juniperinus (Nalepa) sensu Keifer (Acari: Eriophyoidea: Phytoptidae) causes irregular development of buds, shoot deformations and stunted growth of trees, resulting in a serious threat to nurseries and young stands of Cupressus sempervirens L. (Mediterranean cypress). Recently, some cypress clones selected for their resistance to the fungal canker agent Seiridium cardinale (Wag.) have shown high susceptibility to the mite. Considering its tiny body, its hidden lifestyle inside the buds and the probable occurrence of other species (the vagrant Epitrimerus cupressi (Keifer) is common on the Mediterranean cypress in Italy), detection and monitoring of T. juniperinus require taxonomic expertise and are often time-consuming and challenging before serious damage is discernible. In the present study, a rapid, cost-effective PCR-based method was developed and validated to detect T. juniperinus on cypresses. The cytochrome c oxidase subunit I gene was amplified with degenerate and specific primers, but the latter were the only ones able to discriminate between T. juniperinus and E. cupressi. PCR products distinguished the two species both in a pool of individuals in a mixed population of both species and in single individuals, indicating the sensitivity of the detection method. PCR-RFLP (restriction fragment length polymorphism) by means of XmnI and XbaI endonucleases separated the two species. Furthermore, a washing-sieving protocol was used to make mite collection from the tree sample faster and simpler; this procedure did not interfere with the molecular detection of the species. The possibility of the routine use of this assay to monitor quarantine eriophyoids infesting plant material is discussed. PMID:24030201

  12. Safety and anti-ulcerogenic activity of a novel polyphenol-rich extract of clove buds (Syzygium aromaticum L).

    Science.gov (United States)

    Issac, Abin; Gopakumar, G; Kuttan, Ramadasan; Maliakel, Balu; Krishnakumar, I M

    2015-03-01

    Despite the various reports on the pharmacology of Clove bud [Syzygium aromaticum]-derived essential oil and its major component eugenol, systematic information on the bioactivity of clove polyphenols is very limited. Clove buds being one of the richest sources of dietary polyphenols with many traditional medicinal uses, the present contribution attempted to derive their standardized polyphenol-rich extracts as a water soluble free flowing powder (Clovinol) suitable for functional food applications, without the issues of its characteristic pungency and aroma. The extract was characterized by electrospray ionization-time of flight mass spectrometry (ESI-TOF-MS), and investigated for in vivo antioxidant, anti-inflammatory and anti-ulcerogenic activities. Clovinol showed significant antioxidant and anti-inflammatory effects as measured by cellular antioxidant levels, and the ability to inhibit carrageenan-induced paw swelling in mice. Further investigations revealed its significant anti-ulcerogenic activity (>97% inhibition of ethanol-induced stomach ulcers in Wistar rats when orally administered at 100 mg per kg b.w.) and up regulation of in vivo antioxidants such as superoxide dismutase (SOD), glutathione (GSH), and catalase (CAT). Clovinol also reduced the extent of lipid peroxidation among ulcer induced rats, indicating its usefulness in ameliorating oxidative stress and improving gastrointestinal health, especially upon chronic alcohol consumption. The extract was also shown to be safe and suitable for further investigations and development upon acute toxicity studies at 5 g per kg body weight and 28 days of repeated dose toxicity studies at 2.5 g per kg b.w. PMID:25605463

  13. High-frequency in vitro plantlet regeneration from apical bud as a novel explant of Carum copticum L.

    Institute of Scientific and Technical Information of China (English)

    Mansoureh Salehi; Bahman Hosseini; Zohreh Jabbarzadeh

    2014-01-01

    Objective: To develop an in vitro regeneration system to increase the recovery of Carum copticum L. plantlets as a part of developing a metabolic engineering program.Methods:3-acetic acid and indole butyric acid on direct shoot regeneration and rooting of ajowan from apical bud explants were assessed. All explants were cultured on Murashige and Skoog (MS) medium supplemented with different combinations of 6-benzyl amino purine (BAP) (0, 2.2, 4.4, 8.8µ The efficacy of different concentrations and combinations of 6-benzyladenine, indole-Results: The maximum shoot regeneration frequency (97.5%) and the highest number of shoots produced from apical buds (34 shoots per explant) were obtained on MS medium fortified with BAP (4.4 µmol/L) and IAA (0.5 µmol/L). Low shoot regeneration frequency was observed in BAP free treatments. The effects of different strengths of MS medium and various concentrations of IAA and indole-3- butyric acid on rooting rate, length and average number of roots were also investigated. Application of indole-3- butyric acid (6 µmol/L) in full-strength MS medium, was more effective than IAA and resulted in highest shoot regeneration frequency with the rooting rate of 100% and highest mean number of roots per shoot (41.8). The rooted plantlets were acclimatized successfully in greenhouse conditions with a survival rate of 90%. mol/L) and indole-3-acetic acid (IAA) (0, 0.5, 1.1, 2.2 µmol/L). Conclusion: In this study, a simple and reliable regeneration and acclimatization protocol for Carum copticum has been presented. This protocol can be found very advantageous for a variety of purposes, including mass multiplication of Carum species, medicinal plant breeding studies and transgenic plant production.

  14. Characterization of flower-bud transcriptome and development of genic SSR markers in Asian lotus (Nelumbo nucifera Gaertn..

    Directory of Open Access Journals (Sweden)

    Weiwei Zhang

    Full Text Available Asian lotus (Nelumbo nucifera Gaertn. is the national flower of India, Vietnam, and one of the top ten traditional Chinese flowers. Although lotus is highly valued for its ornamental, economic and cultural uses, genomic information, particularly the expressed sequence based (genic markers is limited. High-throughput transcriptome sequencing provides large amounts of transcriptome data for promoting gene discovery and development of molecular markers.In this study, 68,593 unigenes were assembled from 1.34 million 454 GS-FLX sequence reads of a mixed flower-bud cDNA pool derived from three accessions of N. nucifera. A total of 5,226 SSR loci were identified, and 3,059 primer pairs were designed for marker development. Di-nucleotide repeat motifs were the most abundant type identified with a frequency of 65.2%, followed by tri- (31.7%, tetra- (2.1%, penta- (0.5% and hexa-nucleotide repeats (0.5%. A total of 575 primer pairs were synthesized, of which 514 (89.4% yielded PCR amplification products. In eight Nelumbo accessions, 109 markers were polymorphic. They were used to genotype a sample of 44 accessions representing diverse wild and cultivated genotypes of Nelumbo. The number of alleles per locus varied from 2 to 9 alleles and the polymorphism information content values ranged from 0.6 to 0.9. We performed genetic diversity analysis using 109 polymorphic markers. A UPGMA dendrogram was constructed based on Jaccard's similarity coefficients revealing distinct clusters among the 44 accessions.Deep transcriptome sequencing of lotus flower buds developed 3,059 genic SSRs, making a significant addition to the existing SSR markers in lotus. Among them, 109 polymorphic markers were successfully validated in 44 accessions of Nelumbo. This comprehensive set of genic SSR markers developed in our study will facilitate analyses of genetic diversity, construction of linkage maps, gene mapping, and marker-assisted selection breeding for lotus.

  15. Effects of topical and systemic administration of Eugenia caryophyllata buds essential oil on corneal anesthesia and analgesia.

    Science.gov (United States)

    Khalilzadeh, Emad; Hazrati, Reza; Saiah, Gholamreza Vafaei

    2016-07-01

    Clinical studies suggest that essential oil of Eugenia caryophyllata (Clove) buds (EOEC) is efficacious in the treatment of dental pain. In the present study, we investigated the analgesic and local anesthetic effects of EOEC and its possible mechanisms of action in acute corneal pain in rats. EOEC was extracted by hydro-distillation in a Clevenger type apparatus from clove buds. The acute corneal pain was induced by applying a drop (40 µl) of 5 M NaCl solution on the corneal surface, and the numbers of eye wipes were counted during the first 30 s. The mechanical sensation of the cornea was evaluated by calibrated Von Frey filaments. Systemic administration of EOEC (100 and 200 mg/kg, SC) and morphine (2.5 and 5 mg/kg, IP) produced a significant antinociceptive effect in acute corneal pain. Pretreatment with naloxone or atropine prevented the EOEC-induced analgesia. However, L-arginine and methylene blue did not change the suppressive effect of EOEC on corneal pain response. Topical application of EOEC, eugenol and lidocaine significantly decreased corneal sensitivity. Combination treatments of eugenol (25 µg) with lidocaine (0.5%) and EOEC (50 µg) with lidocaine (0.5%) also significantly suppressed corneal sensitivity. Systemic administration of EOEC produced analgesia in the acute corneal pain through mechanisms that involved both opioidergic and cholinergic systems. In addition, topical instillation of EOEC, eugenol, and lidocaine produced local anesthesia in the rat cornea. Sub-anesthetic doses of EOEC or eugenol produced a significant local anesthetic effect when concurrently used with the sub-anesthetic dose of lidocaine. PMID:27651809

  16. DNA-AuNP networks on cell membranes as a protective barrier to inhibit viral attachment, entry and budding.

    Science.gov (United States)

    Li, Chun Mei; Zheng, Lin Ling; Yang, Xiao Xi; Wan, Xiao Yan; Wu, Wen Bi; Zhen, Shu Jun; Li, Yuan Fang; Luo, Ling Fei; Huang, Cheng Zhi

    2016-01-01

    Viral infections have caused numerous diseases and deaths worldwide. Due to the emergence of new viruses and frequent virus variation, conventional antiviral strategies that directly target viral or cellular proteins are limited because of the specificity, drug resistance and rapid clearance from the human body. Therefore, developing safe and potent antiviral agents with activity against viral infection at multiple points in the viral life cycle remains a major challenge. In this report, we propose a new modality to inhibit viral infection by fabricating DNA conjugated gold nanoparticle (DNA-AuNP) networks on cell membranes as a protective barrier. The DNA-AuNPs networks were found, via a plaque formation assay and viral titers, to have potent antiviral ability and protect host cells from human respiratory syncytial virus (RSV). Confocal immunofluorescence image analysis showed 80 ± 3.8% of viral attachment, 91.1 ± 0.9% of viral entry and 87.9 ± 2.8% of viral budding were inhibited by the DNA-AuNP networks, which were further confirmed by real-time fluorescence imaging of the RSV infection process. The antiviral activity of the networks may be attributed to steric effects, the disruption of membrane glycoproteins and limited fusion of cell membrane bilayers, all of which play important roles in viral infection. Therefore, our results suggest that the DNA-AuNP networks have not only prophylactic effects to inhibit virus attachment and entry, but also therapeutic effects to inhibit viral budding and cell-to-cell spread. More importantly, this proof-of-principle study provides a pathway for the development of a universal, broad-spectrum antiviral therapy.

  17. Fita plástica e fita degradável na enxertia de citros Plastic and degradable tape on citrus budding

    Directory of Open Access Journals (Sweden)

    Roberto Pedroso de Oliveira

    2004-12-01

    Full Text Available Este trabalho foi realizado com o objetivo de avaliar o efeito de fitas plástica e degradável no pegamento e no desenvolvimento de enxertos de citros, em viveiro telado. Borbulhas certificadas de 6-8 mm das cultivares de laranjeira-de-umbigo 'Navelina' (Citrus sinensis (L. Osbeck, tangerineira 'Marisol' (C. reticulata Blanco e do híbrido 'Nova' [C. clementina x (C. paradisi x C. tangerina ] foram enxertadas em limoeiro 'Cravo' (C. limonia Osbeck, com 10 meses de idade, em fevereiro de 2003. A fixação das borbulhas nos porta-enxertos foi feita com fitas de dois tipos: fita plástica de polietileno transparente e fita degradável de parafilme. Os tratamentos foram dispostos em esquema fatorial (3 cultivares x 2 tipos de fita, em delineamento inteiramente ao acaso, com quatro repetições, sendo as unidades experimentais constituídas por dez plantas. A média geral de pegamento dos enxertos foi elevada (99,6%, em razão das condições propícias à enxertia fornecidas no viveiro telado. Não houve efeito do tipo de fita no pegamento dos enxertos nem das cultivares no pegamento e desenvolvimento dos enxertos. No entanto, o desenvolvimento dos enxertos foi sensivelmente maior quando a enxertia foi realizada com fita degradável em relação à fita plástica.The aim of this work was to evaluate the effect of the plastic and degradable tape on budding efficiency and scion development of citrus in nursery under screenhouse. Certificated 6-8 mm buds of the cultivars navel orange 'Navelina' (Citrus sinensis (L. Osbeck, mandarin 'Marisol' (C. reticulata Blanco and the hybrid 'Nova' [C. clementina x (C. paradisi x C. tangerina ] were budded on 10 months rangpur lime (C. limonia Osbeck, in February 2003. The buds were attached to the rootstocks using either one of two types of tape: plastic polyethylene transparent tape and parafilm degradable tape. The experimental design was a factorial entirely randomized with 3 cultivars, 2 tape types and four

  18. Floral Initiation in Response to Planting Date Reveals the Key Role of Floral Meristem Differentiation Prior to Budding in Canola (Brassica napus L.).

    Science.gov (United States)

    Zhang, Yaofeng; Zhang, Dongqing; Yu, Huasheng; Lin, Baogang; Fu, Ying; Hua, Shuijin

    2016-01-01

    In Brassica napus, floral development is a decisive factor in silique formation, and it is influenced by many cultivation practices including planting date. However, the effect of planting date on floral initiation in canola is poorly understood at present. A field experiment was conducted using a split plot design, in which three planting dates (early, 15 September, middle, 1 October, and late, 15 October) served as main plot and five varieties differing in maturity (1358, J22, Zhongshuang 11, Zheshuang 8, and Zheyou 50) employed as subplot. The purpose of this study was to shed light on the process of floral meristem (FM) differentiation, the influence of planting date on growth period (GP) and floral initiation, and silique formation. The main stages of FM developments can be divided into four stages: first, the transition from shoot apical meristem to FM; second, flower initiation; third, gynoecium and androecium differentiation; and fourth, bud formation. Our results showed that all genotypes had increased GPs from sowing to FM differentiation as planting date was delayed while the GPs from FM differentiation to budding varied year by year except the very early variety, 1358. Based on the number of flowers present at the different reproductive stages, the flowers produced from FM differentiation to budding closely approximated the final silique even though the FM differentiated continuously after budding and peaked generally at the middle flowering stage. The ratio of siliques to maximum flower number ranged from 48 to 80%. These results suggest that (1) the period from FM differentiation to budding is vital for effective flower and silique formation although there was no significant correlation between the length of the period and effective flowers and siliques, and (2) the increased number of flowers from budding were generally ineffective. Therefore, maximizing flower numbers prior to budding will improve silique numbers, and reducing FM degeneration should

  19. Floral Initiation in Response to Planting Date Reveals the Key Role of Floral Meristem Differentiation Prior to Budding in Canola (Brassica napus L.)

    Science.gov (United States)

    Zhang, Yaofeng; Zhang, Dongqing; Yu, Huasheng; Lin, Baogang; Fu, Ying; Hua, Shuijin

    2016-01-01

    In Brassica napus, floral development is a decisive factor in silique formation, and it is influenced by many cultivation practices including planting date. However, the effect of planting date on floral initiation in canola is poorly understood at present. A field experiment was conducted using a split plot design, in which three planting dates (early, 15 September, middle, 1 October, and late, 15 October) served as main plot and five varieties differing in maturity (1358, J22, Zhongshuang 11, Zheshuang 8, and Zheyou 50) employed as subplot. The purpose of this study was to shed light on the process of floral meristem (FM) differentiation, the influence of planting date on growth period (GP) and floral initiation, and silique formation. The main stages of FM developments can be divided into four stages: first, the transition from shoot apical meristem to FM; second, flower initiation; third, gynoecium and androecium differentiation; and fourth, bud formation. Our results showed that all genotypes had increased GPs from sowing to FM differentiation as planting date was delayed while the GPs from FM differentiation to budding varied year by year except the very early variety, 1358. Based on the number of flowers present at the different reproductive stages, the flowers produced from FM differentiation to budding closely approximated the final silique even though the FM differentiated continuously after budding and peaked generally at the middle flowering stage. The ratio of siliques to maximum flower number ranged from 48 to 80%. These results suggest that (1) the period from FM differentiation to budding is vital for effective flower and silique formation although there was no significant correlation between the length of the period and effective flowers and siliques, and (2) the increased number of flowers from budding were generally ineffective. Therefore, maximizing flower numbers prior to budding will improve silique numbers, and reducing FM degeneration should

  20. The effects of bud load and regulated deficit irrigation on sugar, organic acid, phenolic compounds and antioxidant activity of Razakı table grape berries

    Directory of Open Access Journals (Sweden)

    Tangolar Semih

    2015-01-01

    Full Text Available This study aims at assessing the effects of increased bud load and irrigation applications on berry quality of the Razakı table grape. Two Regulated Deficit Irrigation (RDI having different irrigation levels (RDI-I and RDI-II based on the growth stages, in addition to a non-irrigated control treatment together with two different bud load practices (K-normal and 2K-two-fold buds of the normal were examined for their effects on quality attributes such as sugar and organic acids contents, phenolic compounds as well as antioxidant capacity of the berries. The non-irrigated vines had highest sugar level (198.86 g/kg in the first year (2013 of the experiment whilst the sugar content of the berries was increased with irrigation (RDI-II in 2014. However the highest organic acid (7.10 g/kg was recorded from the RDI-II treatment in 2013 whereas those of from non-irrigated vines were highest (7.81 g/kg in 2014. Considering the sugar and organic acid content of the berries, bud load effects were not significant. The total phenolic acids were higher under non-irrigated and 2K bud load conditions. Antioxidant activity of berries was increased with RDI-I irrigation and 2K practices in the first year (2013 although no significant effect was recorded in the second year of the experiment. In all applications, glucose among the sugars, tartaric acid among the organic acids, catechin and epicatechin among the phenolic compounds were detected to be higher compared to other components in berries.

  1. A simple model to predict the probability of a peach (Prunus persicae) tree bud to develop as a long or short shoot as a consequence of winter pruning intensity and previous year growth.

    Science.gov (United States)

    Bevacqua, Daniele; Génard, Michel; Lescourret, Françoise

    2012-01-01

    In many woody plants, shoots emerging from buds can develop as short or long shoots. The probability of a bud to develop as a long or short shoot relies upon genetic, environmental and management factors and controlling it is an important issue in commercial orchard. We use peach (Prunus persicae) trees, subjected to different winter pruning levels and monitored for two years, to develop and calibrate a model linking the probability of a bud to develop as a long shoot to winter pruning intensity and previous year vegetative growth. Eventually we show how our model can be used to adjust pruning intensity to obtain a desired proportion of long and short shoots.

  2. Experimental Research on Differentiation-Inducing Growth of Nerve Lateral Bud by HUC-MSCs Chitosan Composite Conduit.

    Science.gov (United States)

    Xiao, Qiang; Zhang, Xuepu; Wu, Yuexin

    2015-11-01

    This study is intended to explore the role of human umbilical-cord-derived mesenchymal stem cells (HUC-MSCs) in nerve end-to-side anastomosis, as well as in the induction and promotion of growth of nerve lateral bud. The chitosan nerve conduit was prepared based on the biological characteristics of chitosan, and the nerve conduit was filled with HUC-MSCs, and was used to bridge the nerve end-to-side anastomotic stoma. The experimental animals were randomly assigned into three groups (10 in each group), and the nerve end-to-side anastomosis was conducted: (1) group A (control group): traditional tibial nerve-common peroneal nerve end-to-side anastomosis; (2) group B (experimental group 1): tibial nerve-common peroneal nerve end-to-side anastomotic stoma bridged with chitosan nerve conduit; (3) group C (experimental group 2): tibial nerve-common peroneal nerve end-to-side anastomotic stoma bridged by chitosan nerve conduit filled with HUC-MSCs. General morphological observation, nerve electrophysiology, and anti-S-100 immunohistochemistry were performed. All experimental animals survived, and no infections were found at operative incisions. The nerve continuity was in good condition through visual observation when sampling, which is mild adhesion to the surrounding tissue and easy to be separated. 12 W HUC-MSCs chitosan composite nerve conduits were degraded completely after operation. Electrophysiological test showed that the nerve conduction velocity (NCV) in group C was significantly higher than that in group A or group B (p fibers and the myelin sheath thickness in group C were larger than those in group A or B. There were no significant differences between the numbers of the medullated fibers and between the myelin sheath thicknesses of groups A and B. By means of anti-S-100 immunohistochemistry, the arrangement of a large number of brown-red proliferating schwann cells around the regenerated nerve fibers in group C could be found, while fewer and sparse brown

  3. Frequent and efficient use of the sister chromatid for DNA double-strand break repair during budding yeast meiosis.

    Directory of Open Access Journals (Sweden)

    Tamara Goldfarb

    Full Text Available Recombination between homologous chromosomes of different parental origin (homologs is necessary for their accurate segregation during meiosis. It has been suggested that meiotic inter-homolog recombination is promoted by a barrier to inter-sister-chromatid recombination, imposed by meiosis-specific components of the chromosome axis. Consistent with this, measures of Holliday junction-containing recombination intermediates (joint molecules [JMs] show a strong bias towards inter-homolog and against inter-sister JMs. However, recombination between sister chromatids also has an important role in meiosis. The genomes of diploid organisms in natural populations are highly polymorphic for insertions and deletions, and meiotic double-strand breaks (DSBs that form within such polymorphic regions must be repaired by inter-sister recombination. Efforts to study inter-sister recombination during meiosis, in particular to determine recombination frequencies and mechanisms, have been constrained by the inability to monitor the products of inter-sister recombination. We present here molecular-level studies of inter-sister recombination during budding yeast meiosis. We examined events initiated by DSBs in regions that lack corresponding sequences on the homolog, and show that these DSBs are efficiently repaired by inter-sister recombination. This occurs with the same timing as inter-homolog recombination, but with reduced (2- to 3-fold yields of JMs. Loss of the meiotic-chromosome-axis-associated kinase Mek1 accelerates inter-sister DSB repair and markedly increases inter-sister JM frequencies. Furthermore, inter-sister JMs formed in mek1Δ mutants are preferentially lost, while inter-homolog JMs are maintained. These findings indicate that inter-sister recombination occurs frequently during budding yeast meiosis, with the possibility that up to one-third of all recombination events occur between sister chromatids. We suggest that a Mek1-dependent reduction in

  4. Conservation of an important montane bamboo Thamnocalamus falconeri, Hook.f. ex Munro through axillary bud proliferation

    Institute of Scientific and Technical Information of China (English)

    Meena Bakshi; Chandrakant Tiwari; Sajad Razvi

    2015-01-01

    Thamnocalamus falconeri, Hook.f. ex Munro., an important bamboo species belonging to the family Po-aceae, locally known as Ringal, occurs in the hills of Ut-tarakhand, India. This species has been traditionally exploited by local communities to support their livelihoods. Increasing needs of the hill villages impose unsustainable pressure on natural stands of Ringal in the Uttarakhand hills and forests have been degraded. The long history of excessive cutting of Ringal from natural forests and the lack of replanting threaten villager livelihoods. Replanting is required to conserve the species. We propose a protocol for generation of planting material through axillary bud proliferation for multiplication and conservation of this species. We collected offsets/rhizomes from a natural stand of T. falconeri in the Chopta Mandal areas (Chamoli dis-trict, India). These were planted at sites of varied elevation and fresh single nodal segments were collected from them as explants. Different sterilization treatments were assessed to combat contamination. Among these, treatment of 0.1%HgCl2 followed by 5% NaOCl, proved best. Among two cytokinin treatments, viz. BAP and Kinetin, singly or in combination, BAP alone (5 mg L-1) proved superior and resulted in 100% bud break. BAP-supplemented MS media yielded maximum vigorous shoot formation (90%) and maximum number of shoots (8.9). Subculturing of shoots on the same medium with similar BAP treatment (5 mg L-1 BAP) enabled continuous production of healthy shoots at similar frequency. Maximum rooting (100%) was recorded on half-strength MS medium supplemented with 5 mg L-1 IBA. Micropropagated plants were hard-ened and acclimatized in soil mixture (2:1:1) and then transplanted to field sites (Magra, Uttarakhand, 1,834 m). Eight to ten months after field transplantation we recorded 100% survival of transplanted material. This microprop-agation protocol could be used successfully for raising a stock of genetically homogenous plant

  5. Comparative Transcriptome Profile of the Cytoplasmic Male Sterile and Fertile Floral Buds of Radish (Raphanus sativus L.).

    Science.gov (United States)

    Mei, Shiyong; Liu, Touming; Wang, Zhiwei

    2016-01-01

    Radish cytoplasmic male sterility (CMS) has been widely used for breeding in Raphanus and Brassica genera. However, the detailed regulation network of the male sterility remains to be determined. Our previous work has shown that the abnormalities in a CMS radish appeared shortly after the tetrad stage when microspores were malformed and the tapetal cells grew abnormally large. In this work, histological analysis shows that anthers are at the tetrad stage when the radish buds are about 1.5 mm in length. Furthermore, a high throughput RNA sequencing technology was employed to characterize the transcriptome of radish buds with length about 1.5 mm from two CMS lines possessing the CMS-inducing orf138 gene and corresponding near-isogenic maintainer lines. A total of 67,140 unigenes were functionally annotated. Functional terms for these genes are significantly enriched in 55 Gene Ontology (GO) groups and 323 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The transcriptome detected transcripts for 72 out of a total of 79 protein genes encoded in the chloroplast genome from radish. In contrast, the radish mitochondrial genome contains 34 protein genes, but only 16 protein transcripts were detected from the transcriptome. The transcriptome comparison between CMS and near-isogenic maintainer lines revealed 539 differentially expressed genes (DEGs), indicating that the false positive rate for comparative transcriptome profiling was clearly decreased using two groups of CMS/maintainer lines with different nuclear background. The level of 127 transcripts was increased and 412 transcripts were decreased in the CMS lines. No change in levels of transcripts except CMS-inducing orf138 was identified from the mitochondrial and chloroplast genomes. Some DEGs which would be associated with the CMS, encoding MYB and bHLH transcription factors, pentatricopeptide repeat (PPR) proteins, heat shock transcription factors (HSFs) and heat shock proteins (HSPs), are discussed. The

  6. Comparative Transcriptome Profile of the Cytoplasmic Male Sterile and Fertile Floral Buds of Radish (Raphanus sativus L.

    Directory of Open Access Journals (Sweden)

    Shiyong Mei

    2016-01-01

    Full Text Available Radish cytoplasmic male sterility (CMS has been widely used for breeding in Raphanus and Brassica genera. However, the detailed regulation network of the male sterility remains to be determined. Our previous work has shown that the abnormalities in a CMS radish appeared shortly after the tetrad stage when microspores were malformed and the tapetal cells grew abnormally large. In this work, histological analysis shows that anthers are at the tetrad stage when the radish buds are about 1.5 mm in length. Furthermore, a high throughput RNA sequencing technology was employed to characterize the transcriptome of radish buds with length about 1.5 mm from two CMS lines possessing the CMS-inducing orf138 gene and corresponding near-isogenic maintainer lines. A total of 67,140 unigenes were functionally annotated. Functional terms for these genes are significantly enriched in 55 Gene Ontology (GO groups and 323 Kyoto Encyclopedia of Genes and Genomes (KEGG pathways. The transcriptome detected transcripts for 72 out of a total of 79 protein genes encoded in the chloroplast genome from radish. In contrast, the radish mitochondrial genome contains 34 protein genes, but only 16 protein transcripts were detected from the transcriptome. The transcriptome comparison between CMS and near-isogenic maintainer lines revealed 539 differentially expressed genes (DEGs, indicating that the false positive rate for comparative transcriptome profiling was clearly decreased using two groups of CMS/maintainer lines with different nuclear background. The level of 127 transcripts was increased and 412 transcripts were decreased in the CMS lines. No change in levels of transcripts except CMS-inducing orf138 was identified from the mitochondrial and chloroplast genomes. Some DEGs which would be associated with the CMS, encoding MYB and bHLH transcription factors, pentatricopeptide repeat (PPR proteins, heat shock transcription factors (HSFs and heat shock proteins (HSPs, are

  7. Relationship between Living Rate of Bud and Emeraence Rate of Ratoon Rice and Characteristics of the First Cropping Mid-season Hybrid Rice

    Institute of Scientific and Technical Information of China (English)

    Lin ZHANG; Hong XIONG; Fuxian XU; Yongchuan ZHU; Xiaoyi GUO; Xingbing ZHOU; Mao LIU

    2012-01-01

    [Objective] The aim was to study the relationship of plant characters with bud living rate and mother stem emergence rate of ratoon rice before harvest. [Method] Eighteen mid-season hybrid rice cultivators approved recently were taken as materials and relationship of plant characters including bud living rate and moth- er stem emergence rate of ratoon rice before harvest was explored, based on relat- ed data. [Result] The shorter rice with lower harvesting index is the main character of high emergence rate for first cropping rice; decline of plant height and increase of ear-bearing percentage would improve emergence of regenerative seedlings. [Conclusion] The research provided scientific reference for breeding of rice cultivars with high regenerative capacity.

  8. The Budding Yeast “Saccharomyces cerevisiae” as a Drug Discovery Tool to Identify Plant-Derived Natural Products with Anti-Proliferative Properties

    Directory of Open Access Journals (Sweden)

    Bouchra Qaddouri

    2011-01-01

    Full Text Available The budding yeast Saccharomyces cerevisiae is a valuable system to study cell-cycle regulation, which is defective in cancer cells. Due to the highly conserved nature of the cell-cycle machinery between yeast and humans, yeast studies are directly relevant to anticancer-drug discovery. The budding yeast is also an excellent model system for identifying and studying antifungal compounds because of the functional conservation of fungal genes. Moreover, yeast studies have also contributed greatly to our understanding of the biological targets and modes of action of bioactive compounds. Understanding the mechanism of action of clinically relevant compounds is essential for the design of improved second-generation molecules. Here we describe our methodology for screening a library of plant-derived natural products in yeast in order to identify and characterize new compounds with anti-proliferative properties.

  9. RA Acts in a Coherent Feed-Forward Mechanism with Tbx5 to Control Limb Bud Induction and Initiation

    Science.gov (United States)

    Nishimoto, Satoko; Wilde, Susan M.; Wood, Sophie; Logan, Malcolm P.O.

    2015-01-01

    Summary The retinoic acid (RA)- and β-catenin-signaling pathways regulate limb bud induction and initiation; however, their mechanisms of action are not understood and have been disputed. We demonstrate that both pathways are essential and that RA and β-catenin/TCF/LEF signaling act cooperatively with Hox gene inputs to directly regulate Tbx5 expression. Furthermore, in contrast to previous models, we show that Tbx5 and Tbx4 expression in forelimb and hindlimb, respectively, are not sufficient for limb outgrowth and that input from RA is required. Collectively, our data indicate that RA signaling and Tbx genes act in a coherent feed-forward loop to regulate Fgf10 expression and, as a result, establish a positive feedback loop of FGF signaling between the limb mesenchyme and ectoderm. Our results incorporate RA-, β-catenin/TCF/LEF-, and FGF-signaling pathways into a regulatory network acting to recruit cells of the embryo flank to become limb precursors. PMID:26212321

  10. RA Acts in a Coherent Feed-Forward Mechanism with Tbx5 to Control Limb Bud Induction and Initiation

    Directory of Open Access Journals (Sweden)

    Satoko Nishimoto

    2015-08-01

    Full Text Available The retinoic acid (RA- and β-catenin-signaling pathways regulate limb bud induction and initiation; however, their mechanisms of action are not understood and have been disputed. We demonstrate that both pathways are essential and that RA and β-catenin/TCF/LEF signaling act cooperatively with Hox gene inputs to directly regulate Tbx5 expression. Furthermore, in contrast to previous models, we show that Tbx5 and Tbx4 expression in forelimb and hindlimb, respectively, are not sufficient for limb outgrowth and that input from RA is required. Collectively, our data indicate that RA signaling and Tbx genes act in a coherent feed-forward loop to regulate Fgf10 expression and, as a result, establish a positive feedback loop of FGF signaling between the limb mesenchyme and ectoderm. Our results incorporate RA-, β-catenin/TCF/LEF-, and FGF-signaling pathways into a regulatory network acting to recruit cells of the embryo flank to become limb precursors.

  11. An in vivo detection system for transient and low-abundant protein interactions and their kinetics in budding yeast.

    Science.gov (United States)

    Brezovich, Andrea; Schuschnig, Martina; Ammerer, Gustav; Kraft, Claudine

    2015-03-01

    Methylation tracking (M-Track) is a protein-proximity assay in Saccharomyces cerevisiae, allowing the detection of transient protein-protein interactions in living cells. The bait protein is fused to a histone lysine methyl transferase and the prey protein to a methylation acceptor peptide derived from histone 3. Upon interaction, the histone 3 fragment is stably methylated on lysine 9 and can be detected by methylation-specific antibodies. Since methylation marking is irreversible in budding yeast and only takes place in living cells, the occurrence of artifacts during cell lysate preparation is greatly reduced, leading to a more accurate representation of native interactions. So far, this method has been limited to highly abundant or overexpressed proteins. However, many proteins of interest are low-abundant, and overexpression of proteins may interfere with their function, leading to an artificial situation. Here we report the generation of a toolbox including a novel cleavage-enrichment system for the analysis of very low-abundant proteins at their native expression levels. In addition, we developed a system for the parallel analysis of two prey proteins in a single cell, as well as an inducible methylation system. The inducible system allows precise control over the time during which the interaction is detected and can be used to determine interaction kinetics. Furthermore, we generated a set of constructs facilitating the cloning-free genomic tagging of proteins at their endogenous locus by homologous recombination, and their expression from centromeric plasmids.

  12. Correlates of Amount Spent on Marijuana Buds During a Discrete Purchase at Medical Marijuana Dispensaries: Results from a Pilot Study.

    Science.gov (United States)

    Kepple, Nancy Jo; Mulholland, Elycia; Freisthler, Bridget; Schaper, Elizabeth

    2016-01-01

    Marijuana purchasing behaviors vary by the purchaser's individual characteristics; however, little is known about patients' purchasing behaviors when buying from medical marijuana dispensaries (MMDs). Our objective was to explore whether patient characteristics were associated with amount spent during one financial transaction at medical marijuana dispensaries. We conducted a pilot study of four purposively sampled MMD locations in Long Beach, California, in 2012. A total of 132 medical marijuana patients (33 patients per dispensary) participated in an exit survey about their demographic characteristics, conditions for their medical marijuana recommendation, amount spent at the dispensary, and cross-streets of where they lived. The sample reported amounts spent on discrete purchases of marijuana buds averaging $40.82 (ranging from $10 to $255). Multivariate regression analyses indicated average amount spent differed significantly by patient age and condition. An increase in 10 years of age was associated with a 10% higher amount spent. Receiving a recommendation for anxiety and/or sleeping problems or other nonspecified conditions was related to higher discrete purchase amounts than chronic pain. This pilot suggests that variations in patient purchasing behaviors from MMDs exist. These purchase behaviors can provide insight into variations in how patients use dispensaries, consume products, and allocate personal resources. PMID:26757234

  13. Positive feedback promotes mitotic exit via the APC/C-Cdh1-separase-Cdc14 axis in budding yeast.

    Science.gov (United States)

    Hatano, Yuhki; Naoki, Koike; Suzuki, Asuka; Ushimaru, Takashi

    2016-10-01

    The mitotic inhibitor securin is degraded via the ubiquitin ligase anaphase-promoting complex/cyclosome (APC/C)-Cdc20 after anaphase onset. This triggers activation of the mitotic protease separase and thereby sister chromatid separation. However, only a proportion of securin molecules are degraded at metaphase-anaphase transition and the remaining molecules are still present in anaphase. The roles of securin and separase in late mitosis remain elusive. Here, we show that securin still inhibits separase to repress mitotic exit in anaphase in budding yeast. APC/C-Cdh1-mediated securin degradation at telophase further liberated separase, which promotes Cdc14 release and mitotic exit. Separase executed these events via its proteolytic action and that in the Cdc14 early release (FEAR) network. Cdc14 release further activated APC/C-Cdh1 in the manner of a positive feedback loop. Thus, the positive feedback promotes mitotic exit via the APC/C-Cdh1-separase-Cdc14 axis. This study shows the importance of the two-step degradation mode of securin and the role of separase in mitotic exit.

  14. Dual control by Cdk1 phosphorylation of the budding yeast APC/C ubiquitin ligase activator Cdh1.

    Science.gov (United States)

    Höckner, Sebastian; Neumann-Arnold, Lea; Seufert, Wolfgang

    2016-07-15

    The antagonism between cyclin-dependent kinases (Cdks) and the ubiquitin ligase APC/C-Cdh1 is central to eukaryotic cell cycle control. APC/C-Cdh1 targets cyclin B and other regulatory proteins for degradation, whereas Cdks disable APC/C-Cdh1 through phosphorylation of the Cdh1 activator protein at multiple sites. Budding yeast Cdh1 carries nine Cdk phosphorylation sites in its N-terminal regulatory domain, most or all of which contribute to inhibition. However, the precise role of individual sites has remained unclear. Here, we report that the Cdk phosphorylation sites of yeast Cdh1 are organized into autonomous subgroups and act through separate mechanisms. Cdk sites 1-3 had no direct effect on the APC/C binding of Cdh1 but inactivated a bipartite nuclear localization sequence (NLS) and thereby controlled the partitioning of Cdh1 between cytoplasm and nucleus. In contrast, Cdk sites 4-9 did not influence the cell cycle-regulated localization of Cdh1 but prevented its binding to the APC/C. Cdk sites 4-9 reside near two recently identified APC/C interaction motifs in a pattern conserved with the human Cdh1 orthologue. Thus a Cdk-inhibited NLS goes along with Cdk-inhibited APC/C binding sites in yeast Cdh1 to relay the negative control by Cdk1 phosphorylation of the ubiquitin ligase APC/C-Cdh1.

  15. Lignan dicarboxylates and terpenoids from the flower buds of Cananga odorata and their inhibitory effects on melanogenesis.

    Science.gov (United States)

    Matsumoto, Takahiro; Nakamura, Seikou; Nakashima, Souichi; Fujimoto, Katsuyoshi; Yoshikawa, Masayuki; Ohta, Tomoe; Ogawa, Keiko; Matsuda, Hisashi

    2014-04-25

    The methanolic extract from the flower buds of Cananga odorata showed an inhibitory effect on melanogenesis in theophylline-stimulated murine B16 melanoma 4A5 cells. From the methanolic extract, two new lignan dicarboxylates, canangalignans I and II, three new terpenoids, canangaterpenes I, II, and III, and eight known compounds were isolated. The structures of these compounds were elucidated on the basis of chemical/physicochemical evidence. Several mono- and sesquiterpene analogues significantly inhibited melanogenesis. In particular, canangaterpene I and (3R,3aR,8aS)-3-isopropyl-8a-methyl-8-oxo-1,2,3,3a,6,7,8,8a-octahydroazulene-5-carbaldehyde exhibited a potent inhibitory effect on melanogenesis [inhibition (%): 34.7±4.2 (p<0.01), 45.5±5.7 (p<0.01) at 1 μM, respectively] without inducing cytotoxicity. Moreover, the biological effect of these compounds was much stronger than that of the reference compound, arbutin. Thus, these isolated terpenoid derivatives may be promising therapeutic agents for the treatment of several skin disorders. PMID:24601675

  16. Anticancer activity of rhamnoallosan against DU-145 cells is kinetically complementary to coexisting Polyphenolics in Psidium guajava budding leaves.

    Science.gov (United States)

    Chen, Kuan-Chou; Hsieh, Chiu-Lan; Huang, Kuan-Dar; Ker, Yaw-Bee; Chyau, Charng-Cherng; Peng, Robert Y

    2009-07-22

    Psidium guajava L. is a valuable farm fruit plant having many medicinal uses. Previously its budding leaves (PE) were shown to contain huge amounts of soluble polyphenolics (SP) including (in mg/g) gallic acid (348), catechin (102), epicatechin (60), rutin (100), quercetin (102), and rutin (100) and to exhibit potent anticancer activity. However, reconstitution of these polyphenolics recovered only 40% of the original bioactivity, and the soluble carbohydrate (SC) portion in PE was suspected to contribute the remaining. PE contained a novel rhamnoallosan, which had a carbohydrate/protein (w/w) ratio = 29.06%/10.27% (=2.83, average molecular mass of 5029 kDa), characteristically evidencing a peptidoglycan, consisting of a composition (mole % ratio) of rhamnose/allose/arabinose/tallose/xylose/fucose/glucose/mannose/galactose = 36.05:24.24:8.76:7.95:7.37:5.90:3.69:3.19:2.85 and of amino acid (in wt %) glycine/leucine/proline/alanine/methionine/isoleucine/valine/histidine/tyrosine/phenylalanine/cysteine/aspartic acid/lysine/glutamic acid = 37.12:12.68:10.05:8.97:5.99:4.89:4.83:4.25:4.05:2.78:1.86:1.10:0.73:0.70. Kinetic analysis showed comparable apparent cell-killing rate coefficients (k(app)) to be 4.03 x 10(3) and 2.92 x 10(3) cells mg(-1) h(-1), respectively, by SP and SC, evidencing the complementary anti-DU-145 bioactivity in nature.

  17. Preformation in vegetative buds of pistachio (Pistacia vera): relationship to shoot morphology, crown structure and rootstock vigor.

    Science.gov (United States)

    Spann, Timothy M; Beede, Robert H; Dejong, Theodore M

    2007-08-01

    Effects of rootstock, shoot carbohydrate status, crop load and crown position on the number of preformed leaf primordia in the dormant terminal and lateral buds of mature and immature 'Kerman' pistachio (Pistacia vera L.) trees were investigated to determine if rootstock vigor is associated with greater shoot preformation. There was no significant variation in preformation related to the factors studied, suggesting strong genetic control of preformation in 'Kerman' pistachio. The growth differences observed among trees on different rootstocks were associated with greater stimulation of neoformed growth in trees on the more vigorous rootstocks. However, most annual extension growth in mature tree crowns was preformed, contrasting with the relatively high rate of neoformation found in young tree crowns. Large amounts of neoformed growth in young trees may allow the trees to become established quickly and secure resources, whereas predominantly preformed growth in mature trees may allow for continued crown expansion without outgrowing available resources. We hypothesized that the stimulation of neoformed growth by the more vigorous rootstocks is associated with greater resource uptake or transport, or both. Understanding the source of variation in shoot extension growth on different rootstocks has important implications for orchard management practices.

  18. Quantitative Determination of Alkaloids in Lotus Flower (Flower Buds of Nelumbo nucifera and Their Melanogenesis Inhibitory Activity

    Directory of Open Access Journals (Sweden)

    Toshio Morikawa

    2016-07-01

    Full Text Available A quantitative analytical method for five aporphine alkaloids, nuciferine (1, nornuciferine (2, N-methylasimilobine (3, asimilobine (4, and pronuciferine (5, and five benzylisoquinoline alkaloids, armepavine (6, norarmepavine (7, N-methylcoclaurine (8, coclaurine (9, and norjuziphine (10, identified as the constituents responsible for the melanogenesis inhibitory activity of the extracts of lotus flowers (the flower buds of Nelumbo nucifera, has been developed using liquid chromatography-mass spectrometry. The optimum conditions for separation and detection of these 10 alkaloids were achieved on a πNAP column, a reversed-phase column with naphthylethyl group-bonded silica packing material, with CH3CN–0.2% aqueous acetic acid as the mobile phase and using mass spectrometry equipped with a positive-mode electrospray ionization source. According to the protocol established, distributions of these 10 alkaloids in the petal, receptacle, and stamen parts, which were separated from the whole flower, were examined. As expected, excellent correlations were observed between the total alkaloid content and melanogenesis inhibitory activity. Among the active alkaloids, nornuciferine (2 was found to give a carbamate salt (2′′ via formation of an unstable carbamic acid (2′ by absorption of carbon dioxide from the air.

  19. Positive feedback promotes mitotic exit via the APC/C-Cdh1-separase-Cdc14 axis in budding yeast.

    Science.gov (United States)

    Hatano, Yuhki; Naoki, Koike; Suzuki, Asuka; Ushimaru, Takashi

    2016-10-01

    The mitotic inhibitor securin is degraded via the ubiquitin ligase anaphase-promoting complex/cyclosome (APC/C)-Cdc20 after anaphase onset. This triggers activation of the mitotic protease separase and thereby sister chromatid separation. However, only a proportion of securin molecules are degraded at metaphase-anaphase transition and the remaining molecules are still present in anaphase. The roles of securin and separase in late mitosis remain elusive. Here, we show that securin still inhibits separase to repress mitotic exit in anaphase in budding yeast. APC/C-Cdh1-mediated securin degradation at telophase further liberated separase, which promotes Cdc14 release and mitotic exit. Separase executed these events via its proteolytic action and that in the Cdc14 early release (FEAR) network. Cdc14 release further activated APC/C-Cdh1 in the manner of a positive feedback loop. Thus, the positive feedback promotes mitotic exit via the APC/C-Cdh1-separase-Cdc14 axis. This study shows the importance of the two-step degradation mode of securin and the role of separase in mitotic exit. PMID:27418100

  20. Scopoletin from the flower buds of Magnolia fargesii inhibits protein glycation, aldose reductase, and cataractogenesis ex vivo.

    Science.gov (United States)

    Lee, Jun; Kim, Nan Hee; Nam, Joo Won; Lee, Yun Mi; Jang, Dae Sik; Kim, Young Sook; Nam, Sang Hae; Seo, Eun-Kyoung; Yang, Min Suk; Kim, Jin Sook

    2010-09-01

    Five compounds previously known structures, scopoletin (1), northalifoline (2), stigmast-4-en-3-one (3), tiliroside (4), and oplopanone (5) were obtained from the flower buds of Magnolia fargesii using chromatographic separation methods. The structures of 1-5 were identified by the interpretation of their spectroscopic data including 1D- and 2D-NMR as well as by comparison with reported values. Three compounds 1-3 were found from M. fargesii for the first time in this study. All the isolates (1-5) were subjected to in vitro bioassays to evaluate the inhibitory activity on advanced glycation end products formation and rat lens aldose reductase (RLAR). Compound 1 showed a remarkable inhibitory activity on advanced glycation end products formation with IC(50) value of 2.93 μM (aminoguanidine: 961 μM), and showed a significant RLAR inhibitory activity with IC(50) value of 22.5 μM (3.3-tetramethyleneglutaric acid: 28.7 μM). Compound 4 exhibited potent inhibitory activity against RLAR (IC(50) = 14.9 μM). In the further experiment ex vivo, cataractogenesis of rat lenses induced with xylose was significantly inhibited by compound 1 treatment.

  1. Detection of QTL for Cold Tolerance at Bud Bursting Stage Using Chromosome Segment Substitution Lines in Rice (Oryza sativa)

    Institute of Scientific and Technical Information of China (English)

    LIN Jing; WANG Yan-ping; WANG Cai-lin; ZHU Wen-yin; ZHANG Ya-dong; ZHU Zhen; ZHAO Ling; CHEN Tao; ZHAO Qing-yong; ZHOU Li-hui; FANG Xian-wen

    2011-01-01

    The cold tolerance at the bud bursting stage (CTB) was evaluated at 5℃ by using a set of 95 chromosome segment substitution lines (CSSLs) derived from an indica rice 9311 and a japonica rice Nipponbare with a genetic background of 9311.The result showed that six CSSLs had slightly stronger effect on CTB than 9311.Total four quantitative trait loci (QTLs) for CTB were preliminary mapped on chromosomes 5 and 7 by substitution mapping.qCTB-5-1,qCTB-5-2 and qCTB-5-3 were mapped in the region of RM267-RM1237,RM2422-RM6054 and RM3321-RM1054,which were 21.3 cM,27.4 cM and 12.7 cM in genetic distance on rice chromosome 5,respectively.qCTB-7 was mapped in a 6.8-cM region of RM11-RM2752 on rice chromosome 7.

  2. Bud development, flowering and fruit set of Moringa oleifera Lam. (Horseradish Tree as affected by various irrigation levels

    Directory of Open Access Journals (Sweden)

    Quintin Ernst Muhl

    2013-12-01

    Full Text Available Moringa oleifera is becoming increasingly popular as an industrial crop due to its multitude of useful attributes as water purifier, nutritional supplement and biofuel feedstock. Given its tolerance to sub-optimal growing conditions, most of the current and anticipated cultivation areas are in medium to low rainfall areas. This study aimed to assess the effect of various irrigation levels on floral initiation, flowering and fruit set. Three treatments namely, a 900 mm (900IT, 600 mm (600IT and 300 mm (300IT per annum irrigation treatment were administered through drip irrigation, simulating three total annual rainfall amounts. Individual inflorescences from each treatment were tagged during floral initiation and monitored throughout until fruit set. Flower bud initiation was highest at the 300IT and lowest at the 900IT for two consecutive growing seasons. Fruit set on the other hand, decreased with the decrease in irrigation treatment. Floral abortion, reduced pollen viability as well as moisture stress in the style were contributing factors to the reduction in fruiting/yield observed at the 300IT. Moderate water stress prior to floral initiation could stimulate flower initiation, however, this should be followed by sufficient irrigation to ensure good pollination, fruit set and yield.

  3. Preformation in vegetative buds of pistachio (Pistacia vera): relationship to shoot morphology, crown structure and rootstock vigor.

    Science.gov (United States)

    Spann, Timothy M; Beede, Robert H; Dejong, Theodore M

    2007-08-01

    Effects of rootstock, shoot carbohydrate status, crop load and crown position on the number of preformed leaf primordia in the dormant terminal and lateral buds of mature and immature 'Kerman' pistachio (Pistacia vera L.) trees were investigated to determine if rootstock vigor is associated with greater shoot preformation. There was no significant variation in preformation related to the factors studied, suggesting strong genetic control of preformation in 'Kerman' pistachio. The growth differences observed among trees on different rootstocks were associated with greater stimulation of neoformed growth in trees on the more vigorous rootstocks. However, most annual extension growth in mature tree crowns was preformed, contrasting with the relatively high rate of neoformation found in young tree crowns. Large amounts of neoformed growth in young trees may allow the trees to become established quickly and secure resources, whereas predominantly preformed growth in mature trees may allow for continued crown expansion without outgrowing available resources. We hypothesized that the stimulation of neoformed growth by the more vigorous rootstocks is associated with greater resource uptake or transport, or both. Understanding the source of variation in shoot extension growth on different rootstocks has important implications for orchard management practices. PMID:17472944

  4. In vitro micropropagation of Desmodium gangeticum (L.) DC (Fam-Fabaceae): a medicinal legume through axillary bud multiplication.

    Science.gov (United States)

    Puhan, Puspashree; Rath, Shiba Prasad

    2012-05-15

    Medicinal plants possess unlimited and untapped wealth of chemical compounds with high drug potential which make these plants useful as sources of biomedicines. The rising demand for herbal medicines in the organized manufacturing sector has ruthlessly exploited the wild growing plant population those have bulk use. So for high rate multiplication of different medicinal plants, it is necessary to standardize the protocol for high regeneration. The efficiency of any regeneration is primarily depends on factors like type of explants used, composition of the medium and type of genotype. Here, we have developed a regeneration protocol of Desmodium gangeticum (L.) DC (Salparni, Fam- Fabaceae) a medicinal plant through axillary bud multiplication. Nodal explants from Desmodium gangeticum plants were cultured on Murashige and Skoog's basal medium with Kn or BA at different concentrations. 0.5 mg L(-1) BA in the medium, showed shoot multiplication. Regenerated shoots measuring 3 cm or more were excised and planted on semi solid basal medium supplemented with varying concentrations of either IAA or IBA for induction of rooting. IBA treatment at 1.0 mg L(-1) was the best eliciting 100% rooting response. The in vitro propagation protocol standardized can be highly useful in raising quality planting materials of Desmodium gangeticum for commercial plantation programmes and germplasm conservation. PMID:24187902

  5. Chemical Composition, Antibacterial Properties and Mechanism of Action of Essential Oil from Clove Buds against Staphylococcus aureus.

    Science.gov (United States)

    Xu, Jian-Guo; Liu, Ting; Hu, Qing-Ping; Cao, Xin-Ming

    2016-01-01

    The essential oil of clove has a wide range of pharmacological and biological activities and is widely used in the medicine, fragrance and flavoring industries. In this work, 22 components of the essential oil obtained from clove buds were identified. Eugenol was the major component (76.23%). The essential oil exhibited strong antibacterial activity against Staphylococcus aureus ATCC 25923 with a minimum inhibitory concentration (MIC) of 0.625 mg/mL, and the antibacterial effects depended on its concentration and action time. Kill-time assays also confirmed the essential oil had a significant effect on the growth rate of surviving S. aureus. We hypothesized that the essential oil may interact with the cell wall and membrane first. On the one hand it destroys cell wall and membranes, next causing the losses of vital intracellular materials, which finally result in the bacterial death. Besides, essential oil penetrates to the cytoplasmic membrane or enters inside the cell after destruction of cell structure, and then inhibits the normal synthesis of DNA and proteins that are required for bacterial growth. These results suggested that the effects of the clove essential oil on the growth inhibition of S. aureus may be at the molecular level rather than only physical damage. PMID:27617990

  6. Chemical Composition, Antibacterial Properties and Mechanism of Action of Essential Oil from Clove Buds against Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Jian-Guo Xu

    2016-09-01

    Full Text Available The essential oil of clove has a wide range of pharmacological and biological activities and is widely used in the medicine, fragrance and flavoring industries. In this work, 22 components of the essential oil obtained from clove buds were identified. Eugenol was the major component (76.23%. The essential oil exhibited strong antibacterial activity against Staphylococcus aureus ATCC 25923 with a minimum inhibitory concentration (MIC of 0.625 mg/mL, and the antibacterial effects depended on its concentration and action time. Kill-time assays also confirmed the essential oil had a significant effect on the growth rate of surviving S. aureus. We hypothesized that the essential oil may interact with the cell wall and membrane first. On the one hand it destroys cell wall and membranes, next causing the losses of vital intracellular materials, which finally result in the bacterial death. Besides, essential oil penetrates to the cytoplasmic membrane or enters inside the cell after destruction of cell structure, and then inhibits the normal synthesis of DNA and proteins that are required for bacterial growth. These results suggested that the effects of the clove essential oil on the growth inhibition of S. aureus may be at the molecular level rather than only physical damage.

  7. The budding yeast nuclear envelope adjacent to the nucleolus serves as a membrane sink during mitotic delay.

    Science.gov (United States)

    Witkin, Keren L; Chong, Yolanda; Shao, Sichen; Webster, Micah T; Lahiri, Sujoy; Walters, Alison D; Lee, Brandon; Koh, Judice L Y; Prinz, William A; Andrews, Brenda J; Cohen-Fix, Orna

    2012-06-19

    The mechanisms that dictate nuclear shape are largely unknown. Here we screened the budding yeast deletion collection for mutants with abnormal nuclear shape. A common phenotype was the appearance of a nuclear extension, particularly in mutants in DNA repair and chromosome segregation genes. Our data suggest that these mutations led to the abnormal nuclear morphology indirectly, by causing a checkpoint-induced cell-cycle delay. Indeed, delaying cells in mitosis by other means also led to the appearance of nuclear extensions, whereas inactivating the DNA damage checkpoint pathway in a DNA repair mutant reduced the fraction of cells with nuclear extensions. Formation of a nuclear extension was specific to a mitotic delay, because cells arrested in S or G2 had round nuclei. Moreover, the nuclear extension always coincided with the nucleolus, while the morphology of the DNA mass remained largely unchanged. Finally, we found that phospholipid synthesis continued unperturbed when cells delayed in mitosis, and inhibiting phospholipid synthesis abolished the formation of nuclear extensions. Our data suggest a mechanism that promotes nuclear envelope expansion during mitosis. When mitotic progression is delayed, cells sequester the added membrane to the nuclear envelope associated with the nucleolus, possibly to avoid disruption of intranuclear organization.

  8. Immunolocalization of Arabinogalactan Proteins and Pectins in Floral Buds of Cucumber (Cucumis sativus L.) During Sex Determination

    Institute of Scientific and Technical Information of China (English)

    Yi-Ben PENG; Cheng ZOU; Hua-Qin GONG; Shu-Nong BAI; Zhi-Hong XU; Yi-Qin LI

    2005-01-01

    Arabinogalactan proteins (AGPs) and pectins were detected in the floral buds of cucumber (Cucumis sativus L.) during its sex determination using the following monoclonal antibodies: MAC 207 (recognizes AGP epitopes); JIM 8 (recognizes a subset ofAGP epitopes); and JIM 5 and JIM 7 (epitopes of pectins esterified to various degrees). In the stem apex meristem (SAM) of the cucumber, epitopes of MAC 207, JIM 7, and JIM 5 were localized in the cells from second to third peripheral layers when the sex organ primodium began to differentiate; epitopes of MAC 207 and JIM 5 were also detected in the ragged edge cells. A very dense labeling signal with MAC 207 was observed in the carpel and pistil primodium. The AGP epitopes recognized by JIM 8 were localized in the anther of the male flower and the anther-like portion of the stagnant stamen of the female flower. This suggests that the AGPs and pectins in the SAM of the cucumber are closely associated with the differentiation of the SAM, from meristematic cells to floral primodium. The subset of AGPs recognized by JIM 8 may play an important role in stamen formation.

  9. Ultrasound-assisted extraction and quantitation of oils from Syzygium aromaticum flower bud (clove) with supercritical carbon dioxide.

    Science.gov (United States)

    Yang, Yu-Chiao; Wei, Ming-Chi; Hong, Show-Jen

    2014-01-01

    This study evaluated ultrasound-assisted supercritical carbon dioxide (USC-CO2) extraction for determining the extraction yields of oils and the contents of eugenol, β-caryophyllene, eugenyl acetate and α-humulene from clove buds. Compared to traditional SC-CO2 extraction, USC-CO2 extraction might provide a 13.5% increase in the extraction yield for the oil while utilizing less severe operating parameters, such as temperature, pressure, CO2 flow rate and the time consumed by the process. Our results were comparable to those obtained using the heat reflux extraction method, though the yield was improved by 20.8% using USC-CO2. In kinetic studies, the USC-CO2 extraction of clove oil followed second-order kinetics. The activation energy for the oil extraction was 76.56kJ/mol. The USC-CO2 procedure facilitated the use of mild extraction conditions, improved extraction efficiency and the quality of products and is a potential method for industry. PMID:24290173

  10. Self-emulsification of alkaline-dissolved clove bud oil by whey protein, gum arabic, lecithin, and their combinations.

    Science.gov (United States)

    Luo, Yangchao; Zhang, Yue; Pan, Kang; Critzer, Faith; Davidson, P Michael; Zhong, Qixin

    2014-05-14

    Low-cost emulsification technologies using food ingredients are critical to various applications. In the present study, a novel self-emulsification technique was studied to prepare clove bud oil (CBO) emulsions, without specialized equipment or organic solvents. CBO was first dissolved in hot alkaline solutions, added at 1% v/v into neutral solutions with 1% w/v emulsifier composed of whey protein concentrate (WPC), gum arabic, lecithin, or their equal mass mixtures, and adjusted to pH 7.0. The self-emulsification process did not affect UV-vis absorption spectrum, reversed-phase HPLC chromatogram, or antimicrobial activity of CBO against Escherichia coli O157:H7, Listeria monocytogenes Scott A, and Salmonella Enteritidis. The entrapment efficiency after extraction by petroleum ether was determined to be about 80%. Most emulsions were stable during 7 days of storage. Emulsions prepared with WPC had smaller particles, whereas emulsions prepared with emulsifier mixtures had more stable particle dimensions. The studied self-emulsification technique may find numerous applications in the preparation of low-cost food emulsions. PMID:24758517

  11. Three Different Pathways Prevent Chromosome Segregation in the Presence of DNA Damage or Replication Stress in Budding Yeast.

    Directory of Open Access Journals (Sweden)

    Gloria Palou

    2015-09-01

    Full Text Available A surveillance mechanism, the S phase checkpoint, blocks progression into mitosis in response to DNA damage and replication stress. Segregation of damaged or incompletely replicated chromosomes results in genomic instability. In humans, the S phase checkpoint has been shown to constitute an anti-cancer barrier. Inhibition of mitotic cyclin dependent kinase (M-CDK activity by Wee1 kinases is critical to block mitosis in some organisms. However, such mechanism is dispensable in the response to genotoxic stress in the model eukaryotic organism Saccharomyces cerevisiae. We show here that the Wee1 ortholog Swe1 does indeed inhibit M-CDK activity and chromosome segregation in response to genotoxic insults. Swe1 dispensability in budding yeast is the result of a redundant control of M-CDK activity by the checkpoint kinase Rad53. In addition, our results indicate that Swe1 is an effector of the checkpoint central kinase Mec1. When checkpoint control on M-CDK and on Pds1/securin stabilization are abrogated, cells undergo aberrant chromosome segregation.

  12. Three Different Pathways Prevent Chromosome Segregation in the Presence of DNA Damage or Replication Stress in Budding Yeast.

    Science.gov (United States)

    Palou, Gloria; Palou, Roger; Zeng, Fanli; Vashisht, Ajay A; Wohlschlegel, James A; Quintana, David G

    2015-09-01

    A surveillance mechanism, the S phase checkpoint, blocks progression into mitosis in response to DNA damage and replication stress. Segregation of damaged or incompletely replicated chromosomes results in genomic instability. In humans, the S phase checkpoint has been shown to constitute an anti-cancer barrier. Inhibition of mitotic cyclin dependent kinase (M-CDK) activity by Wee1 kinases is critical to block mitosis in some organisms. However, such mechanism is dispensable in the response to genotoxic stress in the model eukaryotic organism Saccharomyces cerevisiae. We show here that the Wee1 ortholog Swe1 does indeed inhibit M-CDK activity and chromosome segregation in response to genotoxic insults. Swe1 dispensability in budding yeast is the result of a redundant control of M-CDK activity by the checkpoint kinase Rad53. In addition, our results indicate that Swe1 is an effector of the checkpoint central kinase Mec1. When checkpoint control on M-CDK and on Pds1/securin stabilization are abrogated, cells undergo aberrant chromosome segregation. PMID:26332045

  13. Marketingová strategie Gymnázia olympijských nadějí České Budějovice

    OpenAIRE

    Lundáková, Kateřina

    2010-01-01

    The main topic of this thesis is marketing communications of Gymnasium Olympic hopes in České Budějovice. The first part deals with the theory, which is focused on school communication mix, the school's image and marketing strategy. Then this thesis present actual marketing activities of school, describes the SWOT analysis and shows realisation of questionnaire survey and its post evaluation. Heart of the thesis is proposal of specific marketing strategy for GON, which is based on the results...

  14. Komparace marketingových strategií SK Dynamo České Budějovice a FK Olympie Týn nad Vltavou

    OpenAIRE

    Brom, Josef

    2014-01-01

    This diploma thesis deals with sport marketing - specifically with marketing strategies comparison of professional and amateur football clubs. Forms of commercial communications are described and specifics of sport marketing as well, then there SK Dynamo České Budějovice and FK Olympie Týn nad Vltavou presented including their marketing strategies. In frame of this work there was performed survey focused on perception of the club among it's own fans. The results of the survey were also compar...

  15. Zhodnocení finanční situace Dopravního podniku města České Budějovice, a. s.

    OpenAIRE

    Beránková, Monika

    2016-01-01

    This bachelor thesis investigates the financial situation of the Public Transport Company in České Budějovice, a. s. between 2005 and 2014 and the goal of this thesis is on the basis of date from accounting statement to suggest appropriate methods to assess financial situation of company and apply these methods. In the thesis are calculated horizontal and vertical analysis of the accounting statement, analysis of financial ratio, operating ratios and net working capital. Results of the Public...

  16. Flower-bud formation in explants of photoperiodic and day-neutral Nicotiana biotypes and its bearing on the regulation of flower formation

    Energy Technology Data Exchange (ETDEWEB)

    Rajeevan, M.S.; Lang, A. (Michigan State Univ., East Lansing (United States))

    1993-05-15

    The capacity to form flower buds in thin-layer explants was studied in Nicotiana of several species, cultivars, and lines of differing in their response to photoperiod. This capacity was found in all biotypes examined and could extend into sepals and corolla. It varied depending on genotype, source tissue and its developmental state, and composition of the culture medium, particularly the levels of glucose, auxin, and cytokinin. It was greatest in the two day-neutral plants examined, Samsun tobacco and Nicotiana rustica, where it extended from the inflorescence region down the vegetative stem, in a basipetally decreasing gradient; it was least in the two qualitative photoperiodic plants studied, the long-day plant Nicotiana silvestris and the short-day plant Maryland Mammoth tobacco, the quantitative long-day plant Nicotiana alata and the quantitative short-day plant Nicotiana otophora line 38-G-81, where it was limited to the pedicels (and, in some cases, the sepals). Regardless of the photoperiodic response of the source plants, the response was the same in explants cultured under long and short days. The capacity to form flow buds in explants is present in all Nicotiana biotypes studied supports the idea that it is regulated by the same mechanism(s), regardless of the plant's photoperiodic character. However, flower formation in the explants is not identical with de novo flower formation in a hitherto vegetative plant: it is rather the expression of a floral state already established in the plant, although it can vary widely in extent and spatial distribution. Culture conditions that permit flower-bud formation in an explant are conditions that maintain the floral state and encourage its expression; conditions under which no flower buds are formed reduce this state and/or prevent its expression. 14 refs., 5 figs., 3 tabs.

  17. AN INVESTIGATION ON THE BUD-BREAK AND GROWTH OF CUTTINGS OF 420 A AND 5 BB AMERICAN VINE ROOTSTOCKS IRRADIATED WITH DIFFERENT GAMMA DOSES

    OpenAIRE

    Dardeniz, Alper; TAYYAR, Şemun

    2005-01-01

    Viticulture is an important and intensive agricultural branch in Çanakkale province and also in Turkey. Different American vine rootstocks are widely used against phylloxera in our country. Plant breeders have been using gamma radiation for creating new varieties of crops and for obtaining broad genetic diversity for years. In this study, four different doses of gamma rays were applied to 420 A and 5 BB American vine rootstocks when the buds on the cuttings were at the dormant stage. Then, ef...

  18. Možnosti využití hry geocaching v rozvoji cestovního ruchu v ORP České Budějovice

    OpenAIRE

    CEJPEK, Jan

    2016-01-01

    This thesis deals with the potential of geocaching for tourism development in the area of the administrative district of the municipality with extended competence České Budějovice. The main objective is to create own suggestions of geocaching tourist routes along new or already published geocaches in locations where it could help for tourist development. An essential part of the thesis is an analysis of geocaches located in selected district and the usability of these caches for tourism devel...

  19. Ctf3p, the Mis6 budding yeast homolog, interacts with Mcm22p and Mcm16p at the yeast outer kinetochore

    OpenAIRE

    Measday, Vivien; Hailey, Dale W.; Pot, Isabelle; Givan, Scott A.; Hyland, Katherine M.; Cagney, Gerard; Fields, Stan; Davis, Trisha N.; Hieter, Philip

    2002-01-01

    The budding yeast kinetochore is composed of an inner and outer protein complex, which binds to centromere (CEN) DNA and attaches to microtubules. We performed a genetic synthetic dosage lethality screen to identify novel kinetochore proteins in a collection of chromosome transmission fidelity mutants. Our screen identified several new kinetochore-related proteins including YLR381Wp/Ctf3p, which is a member of a conserved family of centromere-binding proteins. Ctf3p interacts with Mcm22p, Mcm...

  20. Lodgepole pine: the first evidence of seed-based somatic embryogenesis and the expression of embryogenesis marker genes in shoot bud cultures of adult trees.

    Science.gov (United States)

    Park, So-Young; Klimaszewska, Krystyna; Park, Ji-Young; Mansfield, Shawn D

    2010-11-01

    Of the various alternatives for cloning elite conifers, somatic embryogenesis (SE) appears to be the best option. In recent years, significant areas of lodgepole pine (Pinus contorta) forest have been devastated by the mountain pine beetle (MPB) in Western Canada. In an attempt to establish an SE propagation system for MPB-resistant lodgepole pine, several families displaying varying levels of resistance were selected for experimentation involving shoot bud and immature seed explants. In bud cultures, eight embryogenic lines were induced from 2 of 15 genotypes following various treatments. Genotype had an important influence on embryogenic culture initiation, and this effect was consistent over time. These lines were identified by microscopic observation and genetic markers. Despite the abundance of early somatic embryos, the cultures have yet to develop into mature embryos. In contrast, immature zygotic embryos (ZEs) cultured from megagametophytes initiated SE at an early dominance stage via nodule-type callus in 1 of 10 genotypes. As part of the study, putative embryogenesis-specific genes, WOX2 (WUSCHELL homeobox 2) and HAP3A, were analyzed in cultures of both shoot bud explants and ZEs. On the basis of these analyses, we postulate that PcHAP3A was expressed mainly in callus and may be involved in cell division, whereas WOX2 was expressed mainly in embryonal mass (EM)-like tissues. The findings from this study, based on molecular assessment, suggest that the cell lines derived from bud cultures were truly EM. Moreover, these experimental observations suggest that PcWOX2 could be used as an early genetic marker to discriminate embryogenic cultures from callus. PMID:20935320

  1. Fusion of nearby inverted repeats by a replication-based mechanism leads to formation of dicentric and acentric chromosomes that cause genome instability in budding yeast

    OpenAIRE

    Paek, Andrew L.; Kaochar, Salma; Jones, Hope; Elezaby, Aly; Shanks, Lisa; Weinert, Ted

    2009-01-01

    Large-scale changes (gross chromosomal rearrangements [GCRs]) are common in genomes, and are often associated with pathological disorders. We report here that a specific pair of nearby inverted repeats in budding yeast fuse to form a dicentric chromosome intermediate, which then rearranges to form a translocation and other GCRs. We next show that fusion of nearby inverted repeats is general; we found that many nearby inverted repeats that are present in the yeast genome also fuse, as does a p...

  2. Effects of Temperature on Root-Forcing and Budding Treatment of Lily Bulbs%温度对百合种球催芽生根的影响

    Institute of Scientific and Technical Information of China (English)

    苗振; 缑辉; 钱振权; 吴红芝

    2014-01-01

    以‘西伯利亚’、‘索邦’和‘木门’3个百合品种种球为试验材料,采用泥炭为基质进行箱栽,种植后把种球置于10℃、12℃和14℃的低温冷库中进行催芽生根处理,以直接栽培于大棚条件下的种球为对照,处理14 d后,测定各处理组的种球鲜质量变化、芽长、茎生根数量、生根率和芽粗。结果表明,12℃下处理‘西伯利亚’、‘索邦’和‘木门’种球种植14 d的生根效果最好,可显著促进茎生根的发生和芽的生长,且整齐度较好。%Lily bulbs ‘Siberia’,‘Sorbonne’,‘Conca D′or’were planted in the plastic box with peat moss, and then treated with 10℃,12℃and 14℃for inducing root and budding.Some bulbs were placed in greenhouse for control.After 14 days,fresh weight,length of buds,number of stem-roots,rooting rates,diameter of buds were measured for analysis.The results showed that stem-roots and buds formation of all three varieties bulbs after trea-ted 14 days under 12℃ were the best,and had high uniformity.

  3. iAID: an improved auxin-inducible degron system for the construction of a 'tight' conditional mutant in the budding yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Tanaka, Seiji; Miyazawa-Onami, Mayumi; Iida, Tetsushi; Araki, Hiroyuki

    2015-08-01

    Isolation of a 'tight' conditional mutant of a gene of interest is an effective way of studying the functions of essential genes. Strategies that use ubiquitin-mediated protein degradation to eliminate the product of a gene of interest, such as heat-inducible degron (td) and auxin-inducible degron (AID), are powerful methods for constructing conditional mutants. However, these methods do not work with some genes. Here, we describe an improved AID system (iAID) for isolating tight conditional mutants in the budding yeast Saccharomyces cerevisiae. In this method, transcriptional repression by the 'Tet-OFF' promoter is combined with proteolytic elimination of the target protein by the AID system. To provide examples, we describe the construction of tight mutants of the replication factors Dpb11 and Mcm10, dpb11-iAID, and mcm10-iAID. Because Dpb11 and Mcm10 are required for the initiation of DNA replication, their tight mutants are unable to enter S phase. This is the case for dpb11-iAID and mcm10-iAID cells after the addition of tetracycline and auxin. Both the 'Tet-OFF' promoter and the AID system have been shown to work in model eukaryotes other than budding yeast. Therefore, the iAID system is not only useful in budding yeast, but also can be applied to other model systems to isolate tight conditional mutants.

  4. Active oxygen metabolism in the floral buds and leaves of the new cytoplasm male sterile(CMS)line and its maintainer line of non-heading Chinese cabbage

    Institute of Scientific and Technical Information of China (English)

    YUAN Jianyu; HOU Xilin; ZHANG Changwei; YE Fan

    2007-01-01

    The cytoplasm male sterile(CMS)line and its maintainer line of the newly-bred non-heading Chinese cabbage were taken as experimental materials.The production rate of superoxide anion radical(O2-),contents of malondialdehyde(MDA)and hydrogen peroxide(H2O2),and the activities of superoxide dismutase(SOD),peroxidase(POD),catalase(CAT)as well as ascorbate peroxidase(APX)were determined in buds of different developmental stages and leaves during vegetative and florescence stages.As showed by the results,there were higher levels of MDA and H2O2 contents,higher 02- production rate and higher activities of SOD,POD,and CAT in the buds of the CMS lines than that in the buds of the fertile lines,while the APX activity was different from the other three enzymes above.There were differences in activities of protective enzymes in leaves but no significant differences in MDA contents and O2- production rate,which showed that the differences in metabolism of active oxygen between CMS and its maintainer were mainly apparent in reproductive organs.

  5. A comparative study of field emission from NanoBuds, nanographite and pure or N-doped single-wall carbon nanotubes

    Energy Technology Data Exchange (ETDEWEB)

    Kleshch, V.I. [Department of Physics, M.V. Lomonosov Moscow State University, 119991 Moscow (Russian Federation); A.M. Prokhorov General Physics Institute, RAS, 119991 Moscow (Russian Federation); Susi, T.; Nasibulin, A.G.; Kauppinen, E.I. [NanoMaterials Group, Department of Applied Physics, Aalto University, P.O. Box 15100, 00076 Aalto, Espoo (Finland); Obraztsova, E.D. [A.M. Prokhorov General Physics Institute, RAS, 119991 Moscow (Russian Federation); Obraztsov, A.N. [Department of Physics, M.V. Lomonosov Moscow State University, 119991 Moscow (Russian Federation); Department of Physics and Mathematics, University of Eastern Finland, 80101 Joensuu (Finland)

    2010-12-15

    Field emission characteristics of multi-emitter flat cathodes prepared from NanoBuds, few-layer graphite flakes (nanographite, NG) and pure or N-doped single-wall carbon nanotubes (SWNTs) have been examined. The cathodes demonstrated a low-voltage electron emission with the threshold field values of few Volts per micron, which were ascribed to the high-aspect ratio of nanocarbons. The films of NanoBuds and NG possess excellent emission pattern homogeneity with an emission site density of more than 10{sup 5} cm{sup -2}. This is much higher than for cathodes made of conventional SWNT films. It was found that the achievable maximal current density depends on the type of carbon material and reaches 10 A/cm{sup 2} for NanoBuds film, while for the NG film cathodes, it is about one order of magnitude less. Possible mechanisms responsible for the experimentally observed differences are discussed. The materials examined are of great interest due to their high-emission characteristics and are promising for the development of new types of vacuum electronic devices. (Copyright copyright 2010 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  6. Ctf3p, the Mis6 budding yeast homolog, interacts with Mcm22p and Mcm16p at the yeast outer kinetochore.

    Science.gov (United States)

    Measday, Vivien; Hailey, Dale W; Pot, Isabelle; Givan, Scott A; Hyland, Katherine M; Cagney, Gerard; Fields, Stan; Davis, Trisha N; Hieter, Philip

    2002-01-01

    The budding yeast kinetochore is composed of an inner and outer protein complex, which binds to centromere (CEN) DNA and attaches to microtubules. We performed a genetic synthetic dosage lethality screen to identify novel kinetochore proteins in a collection of chromosome transmission fidelity mutants. Our screen identified several new kinetochore-related proteins including YLR381Wp/Ctf3p, which is a member of a conserved family of centromere-binding proteins. Ctf3p interacts with Mcm22p, Mcm16p, and the outer kinetochore protein Ctf19p. We used chromatin immunoprecipitation to demonstrate that Ctf3p, Mcm22p, and Mcm16p bind to CEN DNA in a Ctf19p-dependent manner. In addition, Ctf3p, Mcm22p, and Mcm16p have a localization pattern similar to other kinetochore proteins. The fission yeast Ctf3p homolog, Mis6, is required for loading of a CENP-A centromere specific histone, Cnp1, onto centromere DNA. We find however that Ctf3p is not required for loading of the budding yeast CENP-A homolog, Cse4p, onto CEN DNA. In contrast, Ctf3p and Ctf19p fail to bind properly to the centromere in a cse4-1 mutant strain. We conclude that the requirements for CENP-A loading onto centromere DNA differ in fission versus budding yeast. PMID:11782448

  7. Localisation of Abundant and Organ-Specific Genes Expressed in Rosa hybrida Leaves and Flower Buds by Direct In Situ RT-PCR

    Directory of Open Access Journals (Sweden)

    Agata Jedrzejuk

    2012-01-01

    Full Text Available In situ PCR is a technique that allows specific nucleic acid sequences to be detected in individual cells and tissues. In situ PCR and IS-RT-PCR are elegant techniques that can increase both sensitivity and throughput, but they are, at best, only semiquantitative; therefore, it is desirable first to ascertain the expression pattern by conventional means to establish the suitable conditions for each probe. In plants, in situ RT-PCR is widely used in the expression localisation of specific genes, including MADS-box and other function-specific genes or housekeeping genes in floral buds and other organs. This method is especially useful in small organs or during early developmental stages when the separation of particular parts is impossible. In this paper, we compared three different labelling and immunodetection methods by using in situ RT-PCR in Rosa hybrida flower buds and leaves. As target genes, we used the abundant β-actin and RhFUL gene, which is expressed only in the leaves and petals/sepals of flower buds. We used digoxygenin-11-dUTP, biotin-11-dUTP, and fluorescein-12-dUTP-labelled nucleotides and antidig-AP/ streptavidin-fluorescein-labelled antibodies. All of the used methods gave strong, specific signal and all of them may be used in localization of gene expression on tissue level in rose organs.

  8. The budding yeast amphiphysin complex is required for contractile actin ring (CAR assembly and post-contraction GEF-independent accumulation of Rho1-GTP.

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    Michael John Cundell

    Full Text Available The late events of the budding yeast cell division cycle, cytokinesis and cell separation, require the assembly of a contractile actomyosin ring (CAR, primary and secondary septum formation followed by enzymatic degradation of the primary septum. Here we present evidence that demonstrates a role for the budding yeast amphiphysin complex, a heterodimer comprising Rvs167 and Rvs161, in CAR assembly and cell separation. The iqg1-1 allele is synthetically lethal with both rvs167 and rvs161 null mutations. We show that both Iqg1 and the amphiphysin complex are required for CAR assembly in early anaphase but cells are able to complete assembly in late anaphase when these activities are, respectively, either compromised or absent. Amphiphysin dependent CAR assembly is dependent upon the Rvs167 SH3 domain, but this function is insufficient to explain the observed synthetic lethality. Dosage suppression of the iqg1-1 allele demonstrates that endocytosis is required for the default cell separation pathway in the absence of CAR contraction but is unlikely to be required to maintain viability. The amphiphysin complex is required for normal, post-mitotic, localization of Chs3 and the Rho1 GEF, Rom2, which are responsible for secondary septum deposition and the accumulation of GTP bound Rho1 at the bud neck. It is concluded that a failure of polarity establishment in the absence of CAR contraction and amphiphysin function leads to loss of viability as a result of the consequent cell separation defect.

  9. METHODOLOGY FOR FROST-RESISTANCE DETERMINING OF GRAPE VARIETIES (BASED ON EXAMPLE OF STAGE AND CHARACTER OF DAMAGING WINTER BUDS IN GRAPE BUSHES IN JANUARY 2015 IN ZAO “POBEDA” IN TEMRIUK DISTRICT

    OpenAIRE

    Matuzok N. V.; Troshin L. P.; Kulyko I. A.; Druzhkova K. N.

    2015-01-01

    In January 2015, at the Vineyard and ZAO "Pobeda" in Temryuk District of the Krasnodar region, the thermometer dropped to minus 21,6 C, which resulted in significant damage to the central wintering buds. The article presents a method of determining the degree of damage of wintering buds. Based on the results of inspection of variety by sections divided into 3 groups. Group 1: saved between 40 and 72% central. This group includes the following 7 varieties : Pervenets Magaracha (uch. 64) Gibern...

  10. METHODOLOGY FOR FROST-RESISTANCE DETERMINING OF GRAPE VARIETIES (BASED ON EXAMPLE OF STAGE AND CHARACTER OF DAMAGING WINTER BUDS IN GRAPE BUSHES IN JANUARY 2015 IN ZAO “POBEDA” IN TEMRIUK DISTRICT

    Directory of Open Access Journals (Sweden)

    Matuzok N. V.

    2015-02-01

    Full Text Available In January 2015, at the Vineyard and ZAO "Pobeda" in Temryuk District of the Krasnodar region, the thermometer dropped to minus 21,6 C, which resulted in significant damage to the central wintering buds. The article presents a method of determining the degree of damage of wintering buds. Based on the results of inspection of variety by sections divided into 3 groups. Group 1: saved between 40 and 72% central. This group includes the following 7 varieties : Pervenets Magaracha (uch. 64 Gibernal (uch. 74/3, Viorica (uch. 77, Riton (uch. 66, Riesling (uch. 82, Traminer (uch. 73/2, Chardonnay (uch. 73/3. Cutting of bushes should be carried out by the usual way. The task of cutting - compensate the loss of crops due to leaving the greater load. Group 2: Central buds was damaged nearly 70-85%, saved from 20 to 35% of replacement buds. This group includes Merlot (uch. 83, Tsitronyy Magaracha (uch. 69, Augustine (uch. 66, Saperavi (uch. 79, Chardonnay (uch. 74/1, Muscat white (uch. 73 Cabernet Sauvignon (uch. 80. During cutting should be left as much as possible of annual shoots and well-developed secondary shoots. Group 3: damaged more than 85% of central buds, replacement buds significantly damaged . To this group assigned Saperavi (uch. 80, Pinot blanc (uch. 74, Merlot (uch. 80/2, Moldova (uch. 70, Chardonnay (uch. 68, Moldova (uch. 22 Kaberne- Sauvignon (uch. 68, Moldova (uch. 38, Arkadia (uch. 78, Tsimlyansky black (uch. 68. Pre-cutting consists in removing all clearly dead and unusable parts of the bush (dried vines, stumps, annual shoots that will accelerate the development of shoots from the angular buds and resting buds. Considering the high plasticity of grape plants, can not only in accelerated way to restore the crown of bushes damaged by frost, but also get a crop in the current year, using special methods of agricultural technology

  11. Proteção da gema e épocas de forçamento da brotação na enxertia da lima ácida 'Tahiti' Bud protection and timing of bud break treatments on scion, in grafting of Tahiti lime

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    Dierlei dos Santos

    2009-06-01

    Full Text Available A produção de mudas de citros em recipientes é uma tecnologia recente, portanto o sistema de produção encontra-se em aperfeiçoamento, havendo necessidade de informações em várias etapas do sistema produtivo, entre elas os métodos de forçamento da brotação pós-enxertia. Objetivou-se, neste trabalho, estudar o efeito da cobertura das gemas com fitas plásticas e da época de forçamento da brotação sobre o pegamento e desenvolvimento de enxertos da lima ácida 'Tahiti'. O porta-enxerto utilizado foi o limoeiro 'Cravo' (Citrus limonia Osbeck, oriundo da germinação de sementes, e a variedade copa a limeira ácida 'Tahiti' (Citrus latifolia Tanaka. Foram testadas épocas de forçamento de brotação da enxertia (10, 15, 20, 25, 30 e 35 dias após a realização da enxertia e formas de proteção do enxerto (gema coberta e gema exposta. Após a enxertia as plantas foram mantidas em casa de vegetação com esquema de irrigação por gotejamento. O delineamento experimental foi em blocos casualizados, com cinco repetições e 3 plantas por parcela. Não houve interação entre os fatores estudados, sendo que o índice de pegamento dos enxertos não foi influenciado pelos tratamentos e a cobertura total da borbulha pela fita plástica favoreceu a dormência da gema. O forçamento da brotação do enxerto de lima ácida 'Tahiti', utilizando o método de curvamento do porta-enxerto apresentou os melhores resultados quando esse foi realizado aos 10 dias após a enxertia.The production of citrus nursery trees in portable containers is a new developing technology and requires investment in research to acquire information on various stages of their production, including the methods to force post grafting bud break. The aim of this work was to investigate the effect of the coverage of buds with plastic tapes as well as the time to force bud break on the establishment and development of 'Tahiti' lime scions. The rootstock used was the 'Rangpur

  12. Whole-cell imaging of the budding yeast Saccharomyces cerevisiae by high-voltage scanning transmission electron tomography

    Energy Technology Data Exchange (ETDEWEB)

    Murata, Kazuyoshi, E-mail: kazum@nips.ac.jp [National Institute for Physiological Sciences, Okazaki, Aichi 444-8585 (Japan); Esaki, Masatoshi; Ogura, Teru [Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto 860-0811 (Japan); Arai, Shigeo; Yamamoto, Yuta; Tanaka, Nobuo [Ecotopia Science Institute, Nagoya University, Nagoya, Aichi 464-8603 (Japan)

    2014-11-15

    Electron tomography using a high-voltage electron microscope (HVEM) provides three-dimensional information about cellular components in sections thicker than 1 μm, although in bright-field mode image degradation caused by multiple inelastic scattering of transmitted electrons limit the attainable resolution. Scanning transmission electron microscopy (STEM) is believed to give enhanced contrast and resolution compared to conventional transmission electron microscopy (CTEM). Samples up to 1 μm in thickness have been analyzed with an intermediate-voltage electron microscope because inelastic scattering is not a critical limitation, and probe broadening can be minimized. Here, we employed STEM at 1 MeV high-voltage to extend the useful specimen thickness for electron tomography, which we demonstrate by a seamless tomographic reconstruction of a whole, budding Saccharomyces cerevisiae yeast cell, which is ∼3 μm in thickness. High-voltage STEM tomography, especially in the bright-field mode, demonstrated sufficiently enhanced contrast and intensity, compared to CTEM tomography, to permit segmentation of major organelles in the whole cell. STEM imaging also reduced specimen shrinkage during tilt-series acquisition. The fidelity of structural preservation was limited by cytoplasmic extraction, and the spatial resolution was limited by the relatively large convergence angle of the scanning probe. However, the new technique has potential to solve longstanding problems of image blurring in biological specimens beyond 1 μm in thickness, and may facilitate new research in cellular structural biology. - Highlights: • High voltage TEM and STEM tomography were compared to visualize whole yeast cells. • 1-MeV STEM-BF tomography had significant improvements in image contrast and SNR. • 1-MeV STEM tomography showed less specimen shrinkage than the TEM tomography. • KMnO{sub 4} post-treatment permitted segmenting the major cellular components.

  13. Whole-cell imaging of the budding yeast Saccharomyces cerevisiae by high-voltage scanning transmission electron tomography

    International Nuclear Information System (INIS)

    Electron tomography using a high-voltage electron microscope (HVEM) provides three-dimensional information about cellular components in sections thicker than 1 μm, although in bright-field mode image degradation caused by multiple inelastic scattering of transmitted electrons limit the attainable resolution. Scanning transmission electron microscopy (STEM) is believed to give enhanced contrast and resolution compared to conventional transmission electron microscopy (CTEM). Samples up to 1 μm in thickness have been analyzed with an intermediate-voltage electron microscope because inelastic scattering is not a critical limitation, and probe broadening can be minimized. Here, we employed STEM at 1 MeV high-voltage to extend the useful specimen thickness for electron tomography, which we demonstrate by a seamless tomographic reconstruction of a whole, budding Saccharomyces cerevisiae yeast cell, which is ∼3 μm in thickness. High-voltage STEM tomography, especially in the bright-field mode, demonstrated sufficiently enhanced contrast and intensity, compared to CTEM tomography, to permit segmentation of major organelles in the whole cell. STEM imaging also reduced specimen shrinkage during tilt-series acquisition. The fidelity of structural preservation was limited by cytoplasmic extraction, and the spatial resolution was limited by the relatively large convergence angle of the scanning probe. However, the new technique has potential to solve longstanding problems of image blurring in biological specimens beyond 1 μm in thickness, and may facilitate new research in cellular structural biology. - Highlights: • High voltage TEM and STEM tomography were compared to visualize whole yeast cells. • 1-MeV STEM-BF tomography had significant improvements in image contrast and SNR. • 1-MeV STEM tomography showed less specimen shrinkage than the TEM tomography. • KMnO4 post-treatment permitted segmenting the major cellular components

  14. Direct and indirect control of the initiation of meiotic recombination by DNA damage checkpoint mechanisms in budding yeast.

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    Bilge Argunhan

    Full Text Available Meiotic recombination plays an essential role in the proper segregation of chromosomes at meiosis I in many sexually reproducing organisms. Meiotic recombination is initiated by the scheduled formation of genome-wide DNA double-strand breaks (DSBs. The timing of DSB formation is strictly controlled because unscheduled DSB formation is detrimental to genome integrity. Here, we investigated the role of DNA damage checkpoint mechanisms in the control of meiotic DSB formation using budding yeast. By using recombination defective mutants in which meiotic DSBs are not repaired, the effect of DNA damage checkpoint mutations on DSB formation was evaluated. The Tel1 (ATM pathway mainly responds to unresected DSB ends, thus the sae2 mutant background in which DSB ends remain intact was employed. On the other hand, the Mec1 (ATR pathway is primarily used when DSB ends are resected, thus the rad51 dmc1 double mutant background was employed in which highly resected DSBs accumulate. In order to separate the effect caused by unscheduled cell cycle progression, which is often associated with DNA damage checkpoint defects, we also employed the ndt80 mutation which permanently arrests the meiotic cell cycle at prophase I. In the absence of Tel1, DSB formation was reduced in larger chromosomes (IV, VII, II and XI whereas no significant reduction was found in smaller chromosomes (III and VI. On the other hand, the absence of Rad17 (a critical component of the ATR pathway lead to an increase in DSB formation (chromosomes VII and II were tested. We propose that, within prophase I, the Tel1 pathway facilitates DSB formation, especially in bigger chromosomes, while the Mec1 pathway negatively regulates DSB formation. We also identified prophase I exit, which is under the control of the DNA damage checkpoint machinery, to be a critical event associated with down-regulating meiotic DSB formation.

  15. Assessment of knowledge regarding tuberculosis in the context of revised national tuberculosis control program among budding doctors

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    Mausumi Basu

    2014-01-01

    Full Text Available Background: India has the highest burden of tuberculosis (TB in the world with over 2 million incident cases (21% of the global burden. Future physicians should correctly diagnose, treat the cases to prevent the development of newer cases. Materials and Methods: The observational, cross-sectional study was conducted in 2012 among 172 interns. Study tool was an English language, pre-designed, pre-tested, self-administered, semi-structured questionnaire. Study variables were multiple including different aspects of TB knowledge. Then, data were collected from them. Finally, the collected data were tabulated, analyzed and interpreted by proper statistical methods (by percentage. Results: The response rate was 88.4%; mean age was 23.8 (standard deviation ±1.5 years, ranged from 22 to 28 years. A majority of interns (65.4% correctly identified droplet infection as the chief mode of transmission of pulmonary TB (PTB. Again 65.4% interns correctly mentioned coughing for 2 weeks or more as the most important symptom of PTB. Majority (91.0% correctly responded sputum examination for acid fast bacilli for diagnosis. Only a few respondents (25.6% were able to narrate correct categorization of TB. More than 3/4 th (76.9% could correctly name all the five first line anti-TB drugs used in directly observed treatment, short-course (DOTS. Conclusion: The most worrisome finding was the lack of proper knowledge about TB transmission. Poor level of knowledge can prevent the future medical practitioners from suspecting TB. The present study was found comparable with many other study findings. The respondents had moderate knowledge regarding treatment monitoring. There is an urgent need for the huge increase in awareness of DOTS among budding doctors.

  16. High Throughput Analyses of Budding Yeast ARSs Reveal New DNA Elements Capable of Conferring Centromere-Independent Plasmid Propagation.

    Science.gov (United States)

    Hoggard, Timothy; Liachko, Ivan; Burt, Cassaundra; Meikle, Troy; Jiang, Katherine; Craciun, Gheorghe; Dunham, Maitreya J; Fox, Catherine A

    2016-01-01

    The ability of plasmids to propagate in Saccharomyces cerevisiae has been instrumental in defining eukaryotic chromosomal control elements. Stable propagation demands both plasmid replication, which requires a chromosomal replication origin (i.e., an ARS), and plasmid distribution to dividing cells, which requires either a chromosomal centromere for segregation or a plasmid-partitioning element. While our knowledge of yeast ARSs and centromeres is relatively advanced, we know less about chromosomal regions that can function as plasmid partitioning elements. The Rap1 protein-binding site (RAP1) present in transcriptional silencers and telomeres of budding yeast is a known plasmid-partitioning element that functions to anchor a plasmid to the inner nuclear membrane (INM), which in turn facilitates plasmid distribution to daughter cells. This Rap1-dependent INM-anchoring also has an important chromosomal role in higher-order chromosomal structures that enhance transcriptional silencing and telomere stability. Thus, plasmid partitioning can reflect fundamental features of chromosome structure and biology, yet a systematic screen for plasmid partitioning elements has not been reported. Here, we couple deep sequencing with competitive growth experiments of a plasmid library containing thousands of short ARS fragments to identify new plasmid partitioning elements. Competitive growth experiments were performed with libraries that differed only in terms of the presence or absence of a centromere. Comparisons of the behavior of ARS fragments in the two experiments allowed us to identify sequences that were likely to drive plasmid partitioning. In addition to the silencer RAP1 site, we identified 74 new putative plasmid-partitioning motifs predicted to act as binding sites for DNA binding proteins enriched for roles in negative regulation of gene expression and G2/M-phase associated biology. These data expand our knowledge of chromosomal elements that may function in plasmid

  17. Delay in liver bud development during early embryogenesis leads to violation of epicardium formation and its epithelial-to-mesenchymal transformation, but do not affect coronary endothelium formation

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    Pototskaya O. Yu.

    2010-01-01

    Full Text Available Despite intensive investigation of heart embryogenesis the origin of coronary endothelium is still under debate. Existence of close interrelation between proepicardium, liver bud, sinus venosus and early coronary vessels is obvious, but the nature of this interconnection is unclear as well as exact source of endothelial cells. Thus, the purpose of our research was to investigate the effect of inhibition of liver bud development on formation of coronary vessels. To inhibit the liver bud we injected aminoguanidine sulfate into the yolk sack of chick embryos on 14 stage of development by HH (first group - to prevent contact between proepicardium and liver, and on 16 stage (second group to allow contact between proepicardium and liver during a short period of time. In the first group it was observed the violation of epicardium formation and early death during first 3 days of incubation. At embryos it was revealed thinning of myocardium and atrio-ventricular cushions, abnormal looping – presumably due to the absence of epicardium. The death was caused by heart insufficiency. In the second group it was observed the beginning of epicardium formation, but the heart defects were the same as in the first group – presumably due to the absence of epicardium-derived cells. At embryos of this group, who survived till 26 stage of development (4.5 day of incubation, despite absence of cellular component in subepicardial space and almost completely absence of liver, we observed formation of coronary vessels. They were concentrated on the dorsal surface of atrio-vetricular channel and were presented by continuation of endothelium of the sinus venosus.

  18. ALIX Rescues Budding of a Double PTAP/PPEY L-Domain Deletion Mutant of Ebola VP40: A Role for ALIX in Ebola Virus Egress.

    Science.gov (United States)

    Han, Ziying; Madara, Jonathan J; Liu, Yuliang; Liu, Wenbo; Ruthel, Gordon; Freedman, Bruce D; Harty, Ronald N

    2015-10-01

    Ebola (EBOV) is an enveloped, negative-sense RNA virus belonging to the family Filoviridae that causes hemorrhagic fever syndromes with high-mortality rates. To date, there are no licensed vaccines or therapeutics to control EBOV infection and prevent transmission. Consequently, the need to better understand the mechanisms that regulate virus transmission is critical to developing countermeasures. The EBOV VP40 matrix protein plays a central role in late stages of virion assembly and egress, and independent expression of VP40 leads to the production of virus-like particles (VLPs) by a mechanism that accurately mimics budding of live virus. VP40 late (L) budding domains mediate efficient virus-cell separation by recruiting host ESCRT and ESCRT-associated proteins to complete the membrane fission process. L-domains consist of core consensus amino acid motifs including PPxY, P(T/S)AP, and YPx(n)L/I, and EBOV VP40 contains overlapping PPxY and PTAP motifs whose interactions with Nedd4 and Tsg101, respectively, have been characterized extensively. Here, we present data demonstrating for the first time that EBOV VP40 possesses a third L-domain YPx(n)L/I consensus motif that interacts with the ESCRT-III protein Alix. We show that the YPx(n)L/I motif mapping to amino acids 18-26 of EBOV VP40 interacts with the Alix Bro1-V fragment, and that siRNA knockdown of endogenous Alix expression inhibits EBOV VP40 VLP egress. Furthermore, overexpression of Alix Bro1-V rescues VLP production of the budding deficient EBOV VP40 double PTAP/PPEY L-domain deletion mutant to wild-type levels. Together, these findings demonstrate that EBOV VP40 recruits host Alix via a YPx(n)L/I motif that can function as an alternative L-domain to promote virus egress.

  19. The Process Design of Flos Sophorae Buds Dyeing on Wool Fabric%槐米色素对羊毛织物的染色工艺设计

    Institute of Scientific and Technical Information of China (English)

    胥传森

    2011-01-01

    The dyeing property of flos sophorae buds pigments on wool fabric was investigated.On the basis of deeper color depth(K/S),higher color fastness and yellower color(b*),the effects of pH value of dyeing bath,mordant,dyeing temperature and usage of dyestuffs on dyeing performance of flos sophorae buds dyeing on wool fabric were investigated.The results showed that,the optimum dyeing conditions were bath ratio of 1∶30,pH value of 4~6,post-mordant dyeing with aluminum ions,dyeing temperature of 90~98 ℃,the concentration of aluminum mordant of about 5%(owf),and the concentration of the flos sophorae buds dyestuffs 40%~60%(owf).%研究了槐米色素对羊毛织物的染色性能。以表面色深度(K/S)更深、染色牢度更好、颜色更黄(b*)为要求,探讨染液pH值、媒染剂、染色温度及染料用量对槐米染羊毛织物染色性能的影响。研究结果表明,在浴比1∶30条件下,槐米染料对羊毛织物的染色适宜的染液pH值为4~6,宜采用铝后媒染色,且媒染剂的用量为5%(o.w.f)左右,染色温度为90~98℃,染料用量为40%~60%(o.w.f)。

  20. Comparison of glycerolipid biosynthesis in non-green plastids from sycamore (Acer pseudoplatanus) cells and cauliflower (Brassica oleracea) buds.

    Science.gov (United States)

    Alban, C; Joyard, J; Douce, R

    1989-05-01

    The availability of methods to fractionate non-green plastids and to prepare their limiting envelope membranes [Alban, Joyard & Douce (1988) Plant Physiol. 88, 709-717] allowed a detailed analysis of the biosynthesis of lysophosphatidic acid, phosphatidic acid, diacylglycerol and monogalactosyl-diacylglycerol (MGDG) in two different types of non-green starch-containing plastids: plastids isolated from cauliflower buds and amyloplasts isolated from sycamore cells. An enzyme [acyl-ACP (acyl carrier protein):sn-glycerol 3-phosphate acyltransferase) recovered in the soluble fraction of non-green plastids transfers oleic acid from oleoyl-ACP to the sn-1 position of sn-glycerol 3-phosphate to form lysophosphatidic acid. Then a membrane-bound enzyme (acyl-ACP:monoacyl-sn-glycerol 3-phosphate acyltransferase), localized in the envelope membrane, catalyses the acylation of the available sn-2 position of 1-oleoyl-sn-glycerol 3-phosphate by palmitic acid from palmitoyl-ACP. Therefore both the soluble phase and the envelope membranes are necessary for acylation of sn-glycerol 3-phosphate. The major difference between cauliflower (Brassica oleracea) and sycamore (Acer pseudoplatanus) membranes is the very low level of phosphatidate phosphatase activity in sycamore envelope membrane. Therefore, very little diacylglycerol is available for MGDG synthesis in sycamore, compared with cauliflower. These findings are consistent with the similarities and differences described in lipid metabolism of mature chloroplasts from 'C18:3' and 'C16:3' plants (those with MGDG containing C18:3 and C16:3 fatty acids). Sycamore contains only C18 fatty acids in MGDG, and the envelope membranes from sycamore amyloplasts have a low phosphatidate phosphatase activity and therefore the enzymes of the Kornberg-Pricer pathway have a low efficiency of incorporation of sn-glycerol 3-phosphate into MGDG. By contrast, cauliflower contains MGDG with C16:3 fatty acid, and the incorporation of sn-glycerol 3